An English Medium Co. Ed. Senior
Secondary School
Investigatory Project On
SUBM SUBMITTE ITTE B!" B!" SUBMITTE SUBMITTE TO"
Su#hag Singh Sandee$ %ulshesthra &II Sci. B '(.O. Biology)
Mr. Mr.
A*no+ledgement I am over+helmed in all hum#leness and grate,ulness to ac*no+ledge my de$th to all those +ho have hel$ed me to $ut these ideas- +ell a#ove the level o, sim$licity and into something concrete. I +ould li*e to e$ress my s$ecial than*s o, gratitude to my #iology teacher- Mr Mr.. Sandeep Sandee p Kulshesthra Kulshest hra as as +ell as our Princi$al Mrs. Nidhi Bhatia +ho +ho gave gave me the the gold golden en o$$o o$$ort rtun unit ity y to do this this +ond +onder er,u ,ull $roject on the to$ic /A$$lications o, Biotechnology0- +hich also hel$ed me in doing a lot o, research and I came to *no+ a#out so many ne+ things. I am really than*,ul to them. Any attem$t at any level can1t #e satis,actorily com$leted +ithout the su$$ort and guidance o, my Parents and Friends +ho Friends +ho hel$ed me a lot in gathering di2erent in,ormation- collecting data and guiding me ,rom time to time in ma*ing this $roject- des$ite o, their #usy schedules- they gave me di2erent ideas in ma*ing this $roject uni3ue. I am than*,ul to them too. I am ma*ing this $roject not only ,or mar*s #ut to also increase my *no+ledge... Than*ing you Subhag Singh XII Sci. B
Certi4cate This is to certi,y that SUBHAG SINGH o, SINGH o, class XII SCI.B o, GAN !""P SHIKSHA BHA#A$I has success success,ull ,ully y com$le com$leted ted the invest investiga igatory tory $roject on the to$ic %APP&ICA$I'NS APP&ICA$I'NS 'F BI'$"CHN'&'G( under the guidance o, M# . SAN!""P KU&SH"S$H#A '(.O.. Biology) gy) during the session 56789 7897: in the $artial ,ul4lment o, Biology Practical Eamin Eaminati ation on conducted conducted #y C"N$#A& B'A#! 'F S"C'N!A# "!UCA$I'N )AISSC"*. )AISSC"* .
;;;;;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;;;;; Mr. Sandee$ %ulshesthra Mr. Eaminer '(.O. Biology)
Eternal 'C.B.S.E) 'C.B.S.E )
Introduction +hat is Bi,techn,l,g- Bi,techn,l,g- is the use o, living systems and organisms to develo$ or Bi,techn,l,g- is ma*e ma* e $r $roduc oductsts- or
e>ci cient ent in indus dustr trial ial man manu,a u,actu cturin ring g $rocesses.
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Biotech is hel$ing to heal the +orld #y harnessing nature?s o+n tool#o and using our o+n genetic ma*eu$ to heal and guide lines o, research #y"" #y @educing rates o, in,ectious disease Saving millions o, children?s lives Changing the odds o, serious- li,e9threatening conditions a2ecting millions around the +orld Tailoring T ailoring treatme treatments nts to individual individuals s to minimie health ris*s and side e2ects Creating more $recise tools ,or disease detection Com#ating serious illnesses and everyday threats con,ronting the develo$ing +orld.
(istory Throughout the history o, agricultur Throughout agriculturee- ,armer ,armers s have inadverten inadvertently tly altered the genetics o, their cro$s through introducing them to ne+ environments and #reeding them +ith other $lants 9 one o, the 4rst ,orms o, #iotechnology #iotechnology..
These $roc $rocesses esses also +ere included in early /er0entati,n /er0entati,n ,/ ,/ beer beer.. In #re+ing #re+ing-- ma malte lted d gr grain ains s 'c 'cont ontai ainin ning g enymes enymes)) co conv nver ertt st star arch ch ,r ,rom om grains into sugar and then adding s$eci4c yeasts yeasts to to $roduce #eer. In this $rocess- car#ohydrates car#ohydrates in in the grai gr ain ns +er ere e #r #ro o*en do+n into alco al coho hols ls su such ch as et etha hano nol. l. a ate terr other cultures $roduced the $rocess o, lactic acid ,ermentation +h +hic ich h all llo o+e +ed d th the e ,erment ,er mentatio ation n and $r $reser eservati vation on o, other ,orms o, ,ood- such as s,sauce.. er sauce erm men enta tati tion on +a +as s al also so used us ed in th this is ti time me $e $eri riod od to $r $rod oduc uce e lea1 lea1ened ened brea bread d. Alt Althou hough gh the $rocess o, ,ermentation +as not ,ully understood until ouis Pasteur?s Pasteur ?s +or* in 7D8- it is still the 4rst use o, #iotechnology to convert a ,ood source into another ,orm. or th thou ousa sand nds s o, ye year arss- hu huma mans ns ha have ve us used ed se sele lect ctiv ive e #r #ree eedi ding ng to im$r im $rov ove e $r $rod oduct uctio ion n o, cr cro$s o$s an and d liv livest estoc* oc* to use the them m ,or ,oo ,ood. d. In selective #reeding- organisms +ith desira#le characteristics are mated to $roduce o2s$ring +ith the same characteristics. or eam$le- this techni tec hni3u 3ue e +a +as s us used ed +it +ith h co corn rn to $r $rodu oduce ce the lar larges gestt an and d s+ s+eet eetest est cro$s. Biotec Biot echn hnol olog ogy y ha has s al also so le led d to th the e de deve velo lo$m $men entt o, an anti ti#i #iot otic ics. s. In 7F5D- Alean Aleander der leming disc discover overed ed the moul mould d Penicilliu0 Penicilliu0.. (is +or* led to the $uri4cation o, the anti#iotic com$ound ,ormed #y the mould #y (o+ard lorey- Ernst Boris Chain and Gorman (eatley 9 to ,orm +hat +e today *no+ as $enicillin $enicillin.. In 7FH6- $enicillin #ecame availa#le ,or medicinal use to treat #acterial in,ections in humans. The 4eld o, modern #iotechn #iotechnology ology is generally thought o, as having #een #orn in 7F7 +hen Paul Berg?s e$eriments in gene splicing had early ear ly suc succes cess. s. (e (er#e r#ert rt =. Boy Boyer er and Sta Stanl nley ey G. Coh Cohen en sig signi ni4c 4cant antly ly advanced the ne+ technology in 7F5 #y trans,erring genetic material into a #acterium- such that the im$orted material +ould #e re$roduced.
Biotechnology in Agriculture Geneticall- M,di2ed Cr,ps
Geneti Gene tica call ll- 0, 0,di di2e 2ed d cr cr,p ,ps s o orr /M cro$s0 or /#iotech cro$s0 are $lants used in agriculture agriculture-- the GA GA o, o, +hich has #een modi4ed +ith genetic engineering te tech chni ni3u 3ues es.. In mo most st ca case ses s the aim is to introduce a ne+ trait trait to to the $lant +hich does not occur naturally in the th e s$e $ec cies es.. Eam$l $les es in ,ood cro$ o$s s include incl ude res resista istance nce to cer certain tain $est $estss- dise diseases ases-- str stress ess,ul ,ul envi environ ronment mental al conditions- resistance to chemical treatments- reduction o, s$oilage- or im$roving the nutrient $ro4le o, the cro$. Eam$les in non9,ood cro$s incl in clud ude e $r $rod oduc ucti tion on o, $har $harmace maceutic utical al agen agents ts-- #io ,uel elss- an and d other industrially use,ul goods- as +ell as ,or #ioremediation . Plants and cro$s +ith M traits have #een tested more than any other cro$sJ+ith cro$sJ +ith no credi credi#le #le evidence o, harm to humans or animals. In ,actseeds +ith M traits have #een tested more than any other cro$s in the history o, agriculture K +ith no credi#le evidence o, harm to humans or animals. overnmental regulatory agencies- scienti4c organiations and leading health associations +orld+ide agree that ,ood gro+n ,rom GM cr,ps is sa/e sa /e t, ea eatt. Th The e =orl orld d (ea (ealth lth Or Organ gania iatio tionn- the Ame Americ rican an Med Medic ical al Association- the U.S. Gational Academy o, Sciences- the British @oyal Society- among others that have eamined the evidence- all come to the same conclusion" consuming ,oods containing ingredients derived ,rom M cro$s is sa,e to eat and no ris*ier than consuming the same ,oods containing ingredients ,rom cro$ $lants modi4ed #y conventional $lant im$rovement techni3ues. enetic modi4cations have" 7. Made cro$s cro$s more tolerant tolerant to a#iotic stresse stresses s 'col 'coldd- droughtdrought- saltheat). 5. @e @educed duced reliance reliance on chemical chemical $esticides '$est '$est resistant resistant cro$s). cro$s).
L. (el$ed (el$ed to reduce reduce $ost harvest losses losses enhanced enhanced the nutritio nutritional nal value o, the ,ood.
#NA Inter/erence )#NAi* @GA in @GA inte ter, r,er eren ence ce '@ '@GA GAi) i) is a me meth thod od o, #l #loc oc*i *ing ng ge gene ne ,u ,unc ncti tion on #y inserting short se3uences o, ri#onucleic acid '@GA) that match $art o, the target gene1s se3uence- thus no $roteins are $roduced. @GAi has the $otential to #ecome a $o+er,ul thera$eutic a$$roach to+ard targeted and $ersonalied medicine. @GAi has $rovided a +ay to control $ests and diseases- introduce novel $lant traits and increase cro$ yield. Using @GAi @G Ai-- sc scie ient ntis ists ts ha have ve de deve velo lo$e $ed d no nove vell cr cro$ o$s s su such ch as ni nico coti tine ne9, 9,rree to#acc to# accoo- non non9al 9aller lergen genic ic $e $eanu anutsts- dec deca2e a2eina inated ted co co2ee 2ee-- and nut nutri rient ent ,orti4ed maie among many others. Mechanism o, @GA inter,erences as understood is that it comes into $lay +hen a dou#le stranded @GA is introduced either naturally or arti4cially in a cell. An endo ri#onuclease enyme cleaves the long ds@GA into smal sm alll $i $iec eces es o, @G @GA A. Th The e sm smal alll $i $iec eces es co coul uld d #e mi @G @GA A or si @G @GA A de$ending u$on the origin o, long ds@GA i.e. endogenous or eogenous res$ectively. A dou#le stranded @GA may #e generated #y either @GA de$e de $end nden entt @G @GA A $o $oly lyme mera rase se or #idi #idire rectio ctional nal tran transcri scri$tio $tion n o, tra trans$ ns$osa osa#le #le ele elemen ments ts or $hysically introduced. There are several o$$ortunities ,or the a$$l a$ $lic icat atio ions ns o, @G @GAi Ai in cr cro$ o$ scien sc ience ce ,or its im im$r $rov ovem ement ent such as stress tolerance and enhanced nutritional level.This *noc*do+n technology may #e use,ul in indu in duci cing ng ea earl rly y No No+e +eri ring ng-- de dela laye yed d ri ri$e $eni ning ng-- de dela laye yed d se sene nesc scen ence ce-#rea*ing dormancy- stress9,ree $lants- overcoming sel,9sterility- etc . @GA in @GA inte ter, r,er eren ence ce '@ '@GA GAi) i) ha has s rec ecen entl tly y #e #een en de demo mons nstr trat ated ed in $l $lan antt $arasitic nematodes. It is a $otentially $o+er,ul investigative tool ,or the
genome9+ide identi4cation o, gene ,unction that should hel$ im$rove ourr un ou unde ders rsta tand ndin ing g o, $l $lan antt $a $ara rasi siti tic c ne nema mato tode des. s. @G @GAi Ai sh shou ould ld he hel$ l$ iden id enti ti,,y ge gene ne an andd- he henc ncee- $r $rot otei ein n ta tarrge gets ts ,o ,orr ne nema mato tode de co cont ntrrol stra st rate tegi gies es.. Pros os$e $ect cts s ,o ,orr no nove vell res esis ista tanc nce e de de$e $end nd on th the e $l $lan antt generating an e2ective ,orm o, dou#le9stranded @GA in the a#sence o, an end endog ogeno enous us tar target get gen gene e +it +ithou houtt det detrim rimen entt to its itsel, el,.. Th These ese @GA molecules must then #ecome availa#le availa#le to the nematode and #e ca$a#le o, ingestion via its ,eeding tu#e. I, these re3uirements can #e met- cro$ resistance could #e achieved #y a $lant delivering a ds@GA that targets a nematode gene and induces a lethal or highly damaging @GAi e2ect on the $arasite.
Bt t,3in A $rotein that is toic to che+ing insects and is $roduced #y the soil #acterium Bacillus thuringiensis and has long #een used as a #iological pesticide.. By means o, genetic engineering- the genes ,or Bt toin can pesticide #e isolated ,rom Bacillus thuringiensis and trans,erred to $lants. Bacillus thuringiensis )Bt* is )Bt* is a #acteria that $roduces $roteins +hich are toic to insects. But etreme toicity comes at no sur$rise. It1s in the same ,amily o, #acteria as B. anthracis- +hich causes anthra- and B. cereus- +hich causes ,ood $oisoning. The Bt toin toin dissolve in the high $( insect gut and #ecome active. The toins then attac* the gut cells o, the insect- $unching holes in the lining. The Bt s$ores s$ores s$ills out o, the gut and germinate in the insect causing death +ithin a cou$le days. Even though the toin does not *ill the insect immediately- treated $lant $arts $arts +ill +ill not #e damage damaged d #ecau #ecause se the insec insectt sto$s sto$s ,eedin ,eeding g +ithi +ithin n hours. Bt s$or s$ores es do not not s$r s$rea ead d to othe otherr inse insect cts s or caus cause e dise diseas ase e out#rea*s on their o+n.
7. Insect eats Bt crystals crystals and s$ores. 5. The toin #inds to s$eci4c rece$tors in the gut and the insects sto$s eating. L. The crystals cause the gut +all to #re #rea* do+ do+n- allo allo+ +ing ing s$o s$ores and normal gut #acteria to enter the #ody. H. The insect dies as s$ores and gut #acteria $roli,erate in the #ody.
Bt action action is very s$eci4c. i2erent strains o, Bt are are s$eci4c to di2erent rece ece$to $tors in insec sect gut +all. all. Bt toi toici city ty de$e de$end nds s on recog ecogni nii ing ng rece$tors- damage to the gut #y the toin occurs u$on #inding to a rece$tor. Each insect s$ecies $ossesses di2erent ty$es o, rece$tors that +ill match only certain toin $roteins- li*e a loc* to a *ey.
It is #ecause o, this that ,armers have to #e care,ul to match the target $est s$ecies +ith a $articular Bt toin toin $rotein +hich is s$eci4c ,or that insect. This also hel$s the #eni4cal insects #ecause they +ill usually not #e harmed #y that $articular strain o, Bt .
Bt C,tt,n
Bt c, c,tt tt,n ,n is a ge genet netica ically lly mo modi4 di4ed ed or organ ganis ism m 'MO) cotton cotton variety variety+hich $roduces an insecticide insecticide to to #oll+orm #oll+orm.. Strains o, the Bacillus lus thur thuringien ingiensis sis $r #acterium Bacil $rod oduce uce ov over er 566 di di2er 2erent ent Bt to toins ins-each harm,ul to di2erent insects. Most nota#lyBt toins are insecticidal to the larvae o, moths and an d #u #utt tter erNi Nies es-- #eetles #eetles-- co cotto tton n #ol #oll+o l+orrms ms and and ghtu Nies Nies #ut #ut are harmless to other ,orms o, li,e. The gene coding ,or Bt toi toin n has #een inserted into cotton as a transgene transgene-- causing it to $roduce this thi s na natur tural al in insec secti ticid cide e in its tis tissu sues. es. In ma many ny regi re gions ons-- the ma main in $es $ests ts in co comm mmer erci cial al co cotto tton n are le$ido$teran le$ido$teran larvae larvae- +hich are *illed #y the Bt $rotein in thegenetically the genetically modi4ed cotton they cotton they eat at.. Thi his s el elim imiinat ates es th the e nee eed d to us use e lar arge ge amounts o, #road9s$ectrum insecticides to *ill le$ido$teran $ests. This s$ar s$ ares es na natu tura rall in inse sect ct $r $red edat ator ors s in th the e ,a ,arrm ec ecol olog ogy y an and d ,u ,urt rthe herr contri#utes to non insecticide $est management. management . Bt co cott tton on is in ine2 e2ec ecti tive ve ag agai ains nstt ma many ny co cott tton on $e $est sts s su such ch as $lant #ugs-- stin* #ugs#ugs #ugs- and a$hids a$hids de$ending on circumstances it may #e des esir ira a#l #le e to use in ins sec ecti tic cide des s in $r $rev even enttio ion n. A 56 566: 6: stu tud dy do don ne #y Cornell researchersthe Center ,or Chinese Agricultural Policy and the Chinese Academy o, Science on Science on Bt cotton ,arming in Chin Ch ina a ,o ,oun und d th that at a, a,te terr se seve ven n ye year ars s th thes ese e secondary $ests that +ere normally con onttrol ollled #y $es esti tic cide had inc ncrrea ease seddnecessitating the use o, $esticides at similar leve le vels ls to no non9 n9Bt Bt co cott tton on an and d ca caus usin ing g le less ss $ro4 $r o4tt ,or ,ar arm mer ers s #e #ec cau aus se o, th the e et trra e$ense o, M seeds. Mechanism:
Bt cotton +as created through the addition o, genes encoding toin crystals in the Cry grou$ o, endotoin endotoin.. =hen insects attac* and eat the cotton $lant the Cry toins are dissolved due to the high $( level o, the insects stomach. The dissolved and activated Cry molecules #ond to cadherin9li*e $roteins on cells com$rising the #rush #order molecule mole cules. s. The e$it e$itheli helium um o, the #rus #rush h #or #order der mem mem#ran #ranes es se$a se$arate rates s the #ody cavity ,rom the gut +hilst allo+ing access ,or nutrients. The Cry to toin in mo molec lecule ules s att attach ach the themse mselve lves s to s$e s$eci ci4c 4c lo loca catio tions ns on the cadherin9li*e $roteins $resent on the e$ithelial cells o, the midge and
ion channels are ,ormed +hich allo+ the No+ o, $otassium. @egulation o, $otassium concentration is essential and- i, le,t unchec*ed- causes deat de ath h o, ce cell lls. s. u ue e to th the e ,o ,orrma mati tion on o, Cr Cry y io ion n ch chan anne nels ls su su>c >cie ient nt regulation o, $otassium ions is lost and results in the death o, e$ithelial cel ellls. The de dea ath o, such cel ellls crea eattes ga$s in th the e #r #ru ush #or ord der mem#rane.
Advantages: Bt cotton has several advantages over non Bt cotton. The im$ortant advantages o, Bt cotton are #rieNy " •
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Increases yield o, cotton due to e2ective control o, three ty$es o, #oll+orms- vi. American- S$otted and Pin* #oll+orms. Insect Inse cts s #e #elo long nged ed to e$ e$id ido$ o$te tera ra 'B 'Bol oll+ l+or orms ms)) ar are e se sens nsit itiv ive e to crys cr ystal talli line ne en endot doto oic $r $rote otein in $r $rodu oduce ced d #y Bt gen gene e +hi +hich ch in tur turn n $rotects cotton ,rom #oll+orms. @eduction in $esticide use in the cultivation o, Bt cotton in +hich #oll+orms are major $ests. @eduction in the cost o, cultivation and lo+er ,arming ris*s. @educ eduction tion in envi environ ronment mental al $oll $ollutio ution n #y the use o, inse insectic cticides ides rarely. Bt cotton ehi#it genetic resistance or in#uilt resistance +hich is a $erman $er manent ent ty ty$e $e o, re resi sista stanc nce e an and d not a2e a2ect cted ed #y env envir ironm onment ental al ,actors. Thus $rotects cro$ ,rom #oll+orms. Bt co cott tton on is ec eco, o,ri rien endl dly y an and d do does es no nott ha have ve ad adve vers rse e e2 e2ec ectt on $arasites- $redators$redators- #ene4cial insecticides and organi organisms sms $resent in soil.
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It $romotes multi$lication o, $arasites and $redat $redators ors +hich hel$ in controlling the #oll #o ll+o +orrms #y ,e ,eed edin ing g on larvae and eggs o, #oll+orm. Go he heal alth th due to rare insecticides.
haa arrds use o,
Bt cotton are early in maturing as com$ared to non Bt cotton.
Disadvantages: Bt cotton has some limitations •
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(igh cost o, Bt cotton seeds as com$ared to non Bt cotton seeds. E2ecti E2ec tive vene ness ss u$ to 75 756 6 da days ys-- a, a,te terr th that at th the e to toi in n $r $rod oduc ucin ing g e>ciency o, the Bt gene drastically reduces. Ine2ective against suc*ing $ests li*e jassids- a$hids- +hiteNy etc.
Bt cotton in India: Bt cotton is su$$lied in India?s Mah Mahara arasht shtra ra sta state te #y #iotechnology com$any- Mahyco- as the distri#utor.
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The use o, Bt cotton in India ha has s gr gro+ o+n n e$ $on onen enti tial ally ly si sinc nce e it its s introduction. @ecently India has #ecome the num#er one glo#al e$orter o, cotton and the second largest cotton $roducer in the +orld. India has Bikaneri eri Nerm Nerma a and hy#rids such as #red Bt9cotton varieties such as Bikan G((9HH- setting u$ India to #ene4t no+ and +ell into the ,uture. India1s India1 s suc succes cess s has #ee #een n su su#je #ject ct to sc scrut rutin iny y. Monsanto Monsanto?s ?s seed seeds s ar are e e$ensive and lose vigour a,ter one generation- $rom$ting the Indian
Council o, Agricultural @esearch to develo$ a chea$er Bt cotton variety +ith seeds that could #e reused. The cotton incor$orated the cry7Ac gene gen e ,r ,rom om the so soil il #ac #acter teriu ium m Bac Bacil illus lus th thuri uringi ngiens ensis is 'Bt 'Bt))- ma ma*in *ing g the cotton toic to #oll+orms. In $arts o, India cases o, ac3uired resistance against Bt cotton have occurred. The state o, Maharash Maharashtra tra #anned the sale and distri#utio distri#ution n o, Bt cotton in 56 5675 75-- to $r $rom omot ote e lo loca call In Indi dian an se seed edss- +h +hic ich h de dema mand nd le less ss +a +ate terr,ertiliers and $esticide in$ut- #ut li,ted the #an in 567L.
India a$$roved Bt cotton in 5665 no+ it accounts ,or F5 o, all Indian cotton. cott on. Avera verage ge nation+ide nation+ide cotton yields yields +ent ,r ,rom om L65 *gQh *gQha a in the 5665QL season to a $rojected HD7 *gQha in 5677Q75 J u$ 8F.L overall. This chart sho+s the trends in yields- +hich too* o2 a,ter Bt +as introduced in 5665. The gra$hs also sho+ that J and here comes ugly ,actJ in the last H years- as Bt has risen ,rom : to F5 o, India1s cotton- yields have dro$$ed steadily.
Biotechnology in Medicine
Geneticall- "ngineered Insulin )Hu0ulin* Insulin is a $e$tide hormo hormone ne $roduced $roduced #y #eta cells in cells in the $ancreas $ancreas o, various organ or ganis isms ms inc includ ludin ing g hu human man #ei #eings ngs.. It regulates the 0etab,lis0 0etab,lis0 ,/ ,/ carb,h-drates carb,h-drates an an d ,ats #y $r $ro omoti tin ng th the e a#sor or$t $tiion o, glucose ,rom the #lood to s*eletal muscles and muscles and ,at tis tissue sue an and d #y ca caus usin ing g ,at to #e stored rather than used ,or energy. Insulin also inhi#its the $roduction o, glucose #y the liver. Ece$t in the $resence o, the meta#olic disorder dia#etes mellitus and mellitus and meta#olic syndr syndrome ome-- insulin is $rovided +ithin the #ody in a constant $ro$ortion to remove ecess glucose ,rom the #lood- +hich other+ise +ould #e toic. =hen #lood glucose levels ,all #elo+ a certain leve le vell- th the e #o #ody dy #e #egi gins ns to us use e st stor ored ed gl gluc ucos ose e as an en ener ergy gy so sour urce ce through glycogenolysis glycogenolysis-- +hich #rea*s do+n the glycogen stored in the liver and muscles into glucose- +hich can then #e utilied as an energy source. As a central meta#olic control mechanism- its status is also used as a contr control ol signal to other #ody systems 'such as amino acid u$ta*e acid u$ta*e #y #ody cells). In addition- it has several other ana#olic ana#olic e2ects e2ects throug throughout hout the #ody. +hen c,ntr,l ,/ insulin le1els /ails4 diabetes 0ellitus can 0ellitus can result.
Structure: Insulin is com$osed o, t+o di2erent ty$es o, $e$tide chains. Chai Chain n A has has 57 amin amino o acids and Chain B has B has L6 amino acids. Both chains contain al$ha helices #ut no #eta strands. There are L conserved disul2de bridges +hic +hich h hel$ hel$ *ee ee$ $ the the t+o chains together. Insulin can also ,orm dimers in dimers in solution due to the hydrogen #onding #et+een the B chains. The dimers can ,urther int inter erac actt to ,or ,orm heamers due to inter interac actio tion n #et+ee #et+een n hydr hydro$h o$ho#i o#ic c
sur,aces. This scene highlights the highlights the hydro$ho#ic and $olar $arts o, an insulin monomer at a $( o, .
A num# num#er er o, insu insuli lin n vari varian ants ts have have #een #een made ade to ,avo ,avorr eith either er the the monomeric or heameric ,orm. eletion o, the 4ve C terminal residues o, the B chain creates chain creates a monomer only ,orm. This $ortion o, the B chain is involved in hydrogen #onds #et+een #onds #et+een the B chain o, one monomer and the A and B chain o, another monomer .
Need of Genetically Engineered Insulin: The original ,orm o, the +onder cure ,or dia#etesdia#etes- these +ere once the only ty$e o, insulin availa#le- #ut are no+ rarely used. Ani0al insulin +as origin original ally ly made made ,rom ground9u$ animal $ancreas tissue- and then later +as +as etr etrac acte ted d ,rom ,rom healthy animals 'sla 'slaug ught hter ered ed $igs $igs co+s). The meta#olism o, co+s and $igs +as close enough to human meta#olism that their animal insulin also +or*ed +ell in human #odies. Bee, insulin has has L di2e di2errence ences s ,rom ,rom huma human n $or* $or* insu insuli lin n has has 7 di2e di2errence ence ,rom ,rom human. The use o, a miture o, #ee, and $or* insulin +as also $ossi#le. It has #een sho+n that human insulin is less immunogenic than animal insulin. Porcine insulin is most similar to human insulin. The $rimary amino acid se3uences o, #ovine and $orcine insulin di2er ,rom that o, human insulin #y three and one amino acid- res$ectively. This greater dissimilarity #et+een human and #ovine insulin has #een $ostulated to #e the e$la e$lanat nation ion ,or the great greater er antig antigeni enici city ty o, #ovine #ovine insuli insulin n as com$ared +ith $orcine insulin One o, the pr,ble0s 5ith ani0al insulin 5as antib,d- issues. issues . The #ody identi4es them and tries to reject them. Por* insulin di2ers #y 7 amino acid and #ee, insulin #y L amino acids- so the #ody?s immune syst system em can can some someti time mes s recog ecogni nie e them them as ,or ,oreign eign.. Immu Immuno nolo logi gica call com$lications o, insulin thera$y have #een evident since animal insulin #ecam #ecame e avail availa#l a#le e ,or the treat treatmen mentt o, dia#et dia#etes es melli mellitus tus in 7F55. 7F55. In insulin9allergic $atients treated +ith conventional insulin $re$arations-
the insulin9s$eci4c IgE values are o,ten 769 to 569,old higher than in $atients +ithout allergy. It has #een sho+n that human insulin is less immu immuno noge geni nic c than than anim animal al insu insuli lin. n. Por orci cine ne insulin is most similar to human insulin. Cross9 reactivity #et+een human insulin and insulin o, animal origin has #een re$orted. A major $ro# $ro#le lem m is the the cros cross9 s9rrea eact ctiv ivit ity y that that occu occurs rs #et #et+e +een en ant anti9in i9insu sullin anti nti#odies dies and the various animal and human insulin $re$arations in $atients $resenting +ith allergy to animal insulin. The usage o, ani0al insulin has s, greatl- declined in 0,dern ti0es that ti0es that they have largely #een +ithdra+n ,rom the mar*et. Ge+ly diag diagno nose sed d dia# dia#et etic ics s ar are e ty$i ty$ica call lly y give given n synt synthe hesi sie ed d or Geneticall"ngineered hu0an insulin. insulin .
What is !roinsulin"# Pr,insulin is the $rohormone $recu $recurs rsor or to insulin made in the #eta cells o, cells o, the islets islets o, angerh angerhans ans-- s$eci s$ecial alie ied d re regio gions ns o, the $ancreas. $ancreas. Proin oinsulin lin is synthe thesie sied d on mem#rane associated ri#oso ri#osomes mes ,ound ,ound on the ro rough ugh endo$lasmic reticulum- +here it is ,olded and its disul4de #onds are oidi died. ed. It is then hen trans$orted to the olgi a$$aratus +here a$$aratus +here it is $ac*aged int into secr ecreto etory vesi vesic cleses- and +here it is $rocessed #y a series o, $roteases to ,orm mature insulin. insulin. Mature insulin has L8 ,e+er amino acids H are removed altogether- and the remaining L7 ,orm the C9$e$tide. C9$e$tide. The C9$e$tide is a#stracted ,rom the center o, the $roinsulin se3uence the t+o other ends 'the B chain and A chain) remain connected #y disul4de #onds. =hen insulin +as originally $uri4ed ,rom #ovine or #ovine or $orcine $ancreata$orcine $ancreataRLRH RH all the $roinsulin +as not ,ully removed. RL =hen some $eo$le used these insulins- the $roinsulin may have caused the #ody to react +ith a
rash- to resist the insulin- or even to ma*e dents or lum$s in the s*in at the $lac $lace e +her +here e the insulin insulin +as injected injected.. This This can can #e descr descri#e i#ed d as an iatrogenic injury iatrogenic injury due to slight di2erences #et+een the $roinsulin o, di2erent s$ecies. Since the late 7F6s+hen highly $uri4ed $orcine $orcine insulin +as introduced- and the level o, insulin $urity reached reached FF- this ceased to #e a signi4ca signi4cant nt clinical clinical issue. $he 0ain challenge /,r pr,ducti,n ,/ insulin using r!NA techni6ues 5as getting insulin asse0bled int, 0ature /,r0.
$umulin: Hu0ulin +as +as the 4rst 4rst medic medicati ation on $rodu $roduced ced using using moder modern n genet genetic ic engineering techni3ues in +hich actual human GA is inserted into a coli in this host host cell cell 'E. coli this case) case).. Biosy Biosynth ntheti etic c
Chromatogra$hy '(PC) is used to get 766 $ure humulin ,rom the miture o, $roteins. 8. The A and B $oly$e$t $oly$e$tide ide chains chains o, insulin insulin are mied mied together together and conn connec ecte ted d +ith +ith ea each ch othe otherr #y disu disul$ l$hi hide de #on #ond- ,or ,orming ming the the (umulin or synthetic human insulin.
Advantages % Disadvantages of $umulin: (umulin is the one and only human $rotein $roduced in the #acteria +ith identical chemical structure to that o, the natural human insulin. Administration Administration o, humulin reduces the $ossi#ility $ossi#ility o, anti#ody $roduction $roduction and and inNam inNamma mator tory y re res$ s$ons onse e in dia# dia#et etic ic $ati $atien ents ts.. Ma Majo jorr di>culty is the etraction o, humu humuli lin n ,rom ,rom a mit mitur ure e o, host $roteins $resent in the ,ermentation #roth. Go+ days days to over overco come me this this etraction $ro#lem synthetic human human insulin insulin are $roduc $roduced ed in the yeast cell instead o, E. coli using the same $rocedure. As yeast is Eu*aryotes they secrete the +hole humulin molecule +ith $er,ect three dime dimens nsio iona nall stru struct ctur ures es-- reduc educin ing g the the need need ,or ,or com$ com$le le and and time time consuming $uri4cation methods.
Go+ most o, the dia#etic $atients are treated +ith synthetic human insulin. Small grou$ o, $atients claim that e$isodes o, hy$erglycaemic com$l com$lic icati ation ons s have have #een #een incre increase ased d a,ter a,ter shi,t shi,ting ing ,rom ,rom anima animall or origi igin n insulin to humulin. Go study till date sho+s the di2erence #et+een the ,re3uen ,re3uency cy o, hy$ergl hy$erglycae ycaemic mic com$lic com$licatio ations ns in $atient $atient using using humulin humulin 'synthetic human insulin) and animal origin insulin.
Gene $herapGene therap- is therap- is the thera$eutic delivery o, nucleic acid $olymers into $olymers into a $atient?s cells as a drug to trea treatt dise diseas ase. e. ene ene ther thera$ a$y y is an e$erimental techni3ue that uses genes to treat or $revent disease. In the the ,utu ,uturre- this this tec techni3 hni3ue ue may allo+ doctors to treat a disorder #y inserting a gene into a $atient1s cells instead o, usin using g drug drugs s or sur surgery gery. @esearchers are testing severa severall a$$r a$$roac oaches hes to gene gene thera$y- including" •
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@e$la $lacin cing a mutate tated d gene that causes diseas disease e +ith +ith a healt healthy hy co$y o, the gene. InactivatingInactivating- or /*noc*ing out-0 a mutated gene that is ,unctioning im$ro$erly. Introducing a ne+ gene into the #ody to hel$ 4ght a disease.
Although gene thera$y is a $romising treatment o$tion ,or a num#er o, diseas diseases es 'inclu 'includin ding g inheri inherited ted disor disorder derss- some some ty$es ty$es o, cancer cancer-- and and certain viral in,ections)- the techni3ue remains ris*y and is still under study to ma*e sure that it +ill #e sa,e and e2ective. ene thera$y is currently only #eing tested ,or the treatment o, diseases that have no other cures. It should #e noted that not all medical $rocedures that introduce alterations to a $atient?s genetic ma*eu$ can #e considered gene thera$y. thera$y. Bone marro+ marro+ trans$la trans$lantat ntation ion-- and and organ trans$lants trans$lants in in general have #een ,ound to introduce ,oreign GA into $atients. ene thera$y is de4ned #y the $recision o, the $rocedure $rocedure and the intention o, direct thera$eutic e2ects. ene ene ther thera$ a$y y +as +as conc conce$ e$tu tual ali ied ed in 7F5 7F5-- #y auth author ors s +ho +ho urge urged d caution #e,ore commencing human gene thera$y studies. The 4rst attem$t- al#eit an unsuccess,ul unsuccess,ul one- at gene thera$y 'as +ell as the 4rst case o, medical trans,er o, ,oreign genes into humans not counting organ trans$lantation) trans$lantation ) +as $er,ormed #y Martin Cline on 76 uly 7FD6. Cline claimed that one o, the genes in his $atients +as active si si mont months hs late laterr- thou though gh he neve neverr $u#l $u#lis ishe hed d this this data data or had had it
veri4ed and even i, he is correctcorrect- it?s unli*ely unli*ely it $roduced $roduced any signi4cant signi4cant #ene4cial e2ects treating #eta9thalassemia. #eta9thalassemia . The 4rst germ line line gene thera$y consisted o, $roducing a genetically engineered em#ryo in Octo#er 7FF:. The #a#y +as #orn on uly 577FF and +as $roduced #y ta*ing a donor?s egg +ith healthy mitochondria- removing its nuclear GA and GA and 4lling it +ith the nuclear GA GA o, the the #iol #iolog ogic ical al mothe otherr 9 a $roc $roced edur ure e *no+ *no+n n as cyto$lasmic trans,er. trans,er. This $rocedure $rocedure +as re,err re,erred ed to sensationally and and some+hat inaccurately inaccurately in the media as a
The disease is caused #y a mutation in a gene on chromosome 56. A!A de2c de2cie ienc nc- is inhe inheri rite ted d in an aut, aut,s, s,0a 0all rece recess ssi1 i1e e 0anner. 0anner. The gene codes ,or the enyme adenosine deam deamin inas ase e 'A 'AA). A). =itho ithout ut this enyme- the #ody is una#le to #rea* do+n a toic su#stance called deo deoyad yaden enos osin ine. e. The The toi toin n #uilds u$ and destroys in,e in,ect ctio ion9 n94g 4ght htin ing g immu immune ne cells called T and B lym$hoc lym$hocytes ytes.. Because Because AA de4ciency a2ects the immune system- $eo$le +ho have the disorder are more susce$ti#le to all *inds o, in,ections- $articularly those o, the s*in- res$iratory systemand gastrointestinal gastrointestinal tract. They may also #e shorter than normal. normal. Sadlymost #a#ies +ho are #orn +ith the disorder die +ithin a ,e+ months. $reat0ents ,/ A!A !e2cienc- includes" !e2cienc- includes" •
#one marro+ trans$lant gene therap-
•
AA enyme in PE vehicle PE vehicle
'n Septe0ber 8<4 899=4 the 2rst gene therap- therap- to com#at this disease +as $er,ormed $er,ormed #y r. r. =illiam rench Anderson on Anderson on a /,ur>-ear> ,ld ,ld girl girl44 Ashan Ashanti ti !eSil1 !eSil1a a- at the Gati Gation onal al Inst Instit itut utes es o, (eal (ealth th-Bethesda- Maryland- U.S.A.
Conclusion Biotechnology is the ne+ +onder o, science. It is truly multidisci$linary in nature and it encom$asses several disci$lines o, #asic sciences and engineer engineering. ing. The Science Science disci$li disci$lines nes ,rom ,rom +hich +hich #iotechn #iotechnolo ology gy dra+s dra+s heavily are micro#iology- chemistry- #iochemistry- genetics- molecular #iology#iology- immunol immunologyogy- cell cell and tissue tissue cultur culture e and $hysiol $hysiology ogy.. On the engineering side it leans heavily on $rocess chemical and #iochemical engineering since large scale cultivation o, microorganisms and cellstheir do+nstream $rocessing are #ased on them. It c,0es t, us as a great blessing... blessing...
Biotec Biotechn hnolo ology gy utili utilies es the the techni techni3ue 3ue calle called d genetic genetic engin engineer eering ing or recom# recom#inan inantt GA technol technology ogy +here +here a micr microor oorgan ganism ism is isola isolated ted its its gene geneti tic c mate materi rial al is cutcut- mani mani$u $ula late tedd- seal sealed ed-- agai again n inse insert rted ed in an orga or gani nism sm and and allo allo+e +ed d to gro+ gro+ in a suit suita# a#le le envi envirronme onment nt unde underr controlled conditions to get the desired $roduct. It loo*s easy #ut is a very tedious jo# and it ta*es years ,or a research to achieve its goal. i*e every other thing- bi,te bi,tech chn, n,l, l,gg- t,, t,, has has s,0e s,0e har0 har0/u /ull i0pacts" i0pacts" 7. enetic engineering is a very vital $art o, #iotechnology and the cost o, trans,erring genes ,rom one s$ecies to another is very e$ensive- +hich re3uires a huge amount o, ca$ital investment. $he $he c,st c,st ,/ pr,d pr,duc ucin ing g gene geneti tica call ll-> -> 0,di 0,di2e 2ed d plan plants ts and and ani0als are s7-> r,c7eting and r,c7eting and the duration o, return are also not $redicta#le. 5. ene eneti tic c engi engine neer erin ing g cros crosse ses s #oun #ounda dari ries es o, re$r e$roduc oducti tion on #y crossing genes o, s$ecies that are com$letely unrelated hence giving rise to haardous results as +ell as also increasing the ris* o, harming multi$le s$ecies. L. =hen =hen geneti netic c mater terial ,ro ,rom cer erta tain in vir viruses ses is used in the the $roduction o, transgenic cro$s- there are chances that these virus genes +ill com#ine +ith cro$ genes to $roduce more destructive viruses. The consum$tion o, such cro$s is haardous to human health and can cause several li,e9 threatening ailments. ailments. It can also result in cancer- o,ten malignant as +ell. H. Biotec Biotechno hnolog logy y also also $oses $oses a num#e num#err o, envir environ onmen mental tal threat threats. s. eneti enetica cally lly modi modi4es 4es cro$s cro$s o,ten o,ten in,ec in,ectt mona monarc rch h #utter #utteries ies and and other insect s$ecies. The a$$lications a$$lications o, #iotechnology #iotechnology are so #road- and the advantages so com$ com$el elli ling ng-- that that virt virtua uall lly y ever every y indu indust stry ry is usin using g this this tech techno nolo logy gy.. evelo$ments are under+ay in areas as diverse as $harmaceuticalsdiag diagno nost stic icss- tet tetil iles es-- a3ua a3uacu cult ltur uree- ,or ,orestr estryy- chem chemic ical alss- hous househ ehol old d $roducts- environmental cleanu$- ,ood $rocessing and ,orensics to name a ,e+. Biotechnology is ena#ling these industries to ma*e ne+ or #etter $rod $roduc ucts ts-- o,te o,ten n +ith ith grea eate terr s$ee $eed- e>cie >cien ncy and and Nei#il #ility ity. Bi,techn,l,g- 0ust c,ntinue t, be care/ull- regulated s, that the 0a3i0u0 bene2ts are recei1ed 5ith the least ris7 .
Bi#liogra$hy http?@@en.5i7ipedia.,rg@bi,techn,l,ghttp?@@en.5i7ipedia.,rg@bi,techn,l,ghttp?@@en.5i7ipedia.,rg@insulin http?@@en.5i7ipedia.,rg@insulin
http?@@555.gene5atch.,rg@sub>:;: http?@@en.5i7ipedia.,rg@hu0ulin http?@@en.5i7ipedia.,rg@hu0ulin http?@@555.bi,techarticles.c,0@'thers>Article@Hu0an> Insulin>and>#ec,0binant>!NA>$echn,l,g->D=.ht0l https?@@isaaa.,rg@res,urces@p https?@@isaaa.,rg@res,urces@publicati,ns@p,c7 ublicati,ns@p,c7et7@<@de/ault. et7@<@de/ault. asp http?@@555.sciencedirect.c,0@ https?@@en.5i7ipedia.,rg@5i7 https?@@en.5i7ipedia.,rg@5i7i@GeneEtherap i@GeneEtheraphttps?@@en.5i7ipedia.,rg@5i7 https?@@en.5i7ipedia.,rg@5i7i@Aden,sineEd i@Aden,sineEdea0inaseE ea0inaseEde2cie de2cie nchttp?@@555.diabetes.c,.u7@insulin@ani0al>insulin.ht0l Bi,l,g- te3tb,,7 )N.C.".#.$* Class 8;th
Contents • • •
Introduction (istory Biotechnology in Agriculture enetically Modi4ed Cro$s @GA Inter,erence '@GAi) • •
Bt toin Bt cotton Biotechnology in Medicine enetically engineered insulin '(umulin) ene thera$y Conclusion Bi#liogra$hy • •
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