Lecture: Application of Biotechnolgy

unit-4 / Application of biotechnology). The change in the genetic makeup of 6 nganti by nbaticing devireg gene in ( genetic engineering. known as geutic en This allow adv. over the conventional φαν 0 breeding method. new plants & animale have been den Meny new Gusing genche angineerting. The genetically manipulated organisms a are kroven as GMO Geo, are useful in agriculture, industry, medicines ang environment for betterment. human life. negenée plants. plants thal Transgenic plants: The plants. foreign DNA into its (contain foreig as trans genée plants. genonce enonce ans known This plants acquire neco trait and new tralt for few generations. inhext He In some transgenic plants the introduced DNA blocks the normal puncthient of certain. original genes que peants. there are many transgeneic plants cushich have been developed uchth the following properties- y Herblaide resistance place. [ey loung lep reddish bo mecet nesistant of peast tey-BT cotton] (a) anmproved storage proteins. Benvironniental stress resistanes. I flavs, savs tometo. There are diso 9 the quality traits: transgenics cwith eneproved at Golden rice. Genetically modified foods 2) plants weith high rate of photosynthesis. Insect pest Resistant plant: plants that folerat attack. called salted pest Insect & pests are n't plant. Resistant Meny transgente pest resistant, by genetic congineering which manı faniniels + plants are developer deve ars not harmful to plants keep away insects fromicrops of meninise the use of agriculture. chenideal pesticides en of pesticides are sprayed aver plants there is destructions of pesticides by sunlight of they washed diway by pwater which requere are u repeated application for for plant. plant protechich. end hanigenic plant provide protection. through. @BT cotton is such a transgeric all we hich producy Insecticides to against bollworm. The prode production of Ensecticide dess to Bt. introduced into the prent. Et gene ED et gene ara genaa present bacterium Bacillus thekingeness ushich produces endotoxin hekich & bt toxin. This toxin kill welde group of insect om pest but not harmful to mant animals. Hence this gene is isolated 4 Integrated with Tiplasmid of agrobacterium & then transfered to angiosperm Lant. Bt gene from Bacillus thuringene, isolated from ang put unger the control of 35.5 promoter GMVa and introduced into plants. The CaMV promoter obtained from CPT-1 gene of CaMV. The general structure of Bt gene plasmid containing be gene Transcriphon taminahon Signal NED Plys-Accintegrate Tiplaining having Bit gene The method I plasmid bt-gene through Into higher plants like Tobasco, cotton, pea, potato is possible. Advantages:- if mereased yield of crop due to Inbullt resistance to boll worm f other Iweet. Reduction in environmental pollution cost of cultivation due to requced use & perticides. 2lt these transgentes are also scofriendly f doesn't have aquerie effect ons beneficial Ensects eg- predators, parentes etc. Disadvantages:- f High nort i) meffective djatust sucking pests like Jandy Aphids ete if 91 may not be supporting tho growth of o crop varieties growing around it. other • The insect resistant gene of bacillus Bacillus thuringenive bacteria produces a prote wwhich are crystalline in nature and present b melusion in the bacterial spores Q Thés crystal proteins calleg Cay gene as are produced by a gene they produces crystalloid Crystal proteins are responsible for insecticias activity. The Cry genes heich as grouped ento 4 main groups Jiery 1, eng", ery !!! f IV basedon their Insecticidal activity. The very genee code proteins. for either 130 kda or 70 kala @when cry proteins are ingested by insects alkaline juice gi ars dissolved in the alkaline of the Roarjo insect 1 Phey the midgut -The protestes present in the gut of insect process the cay protein hydrolyheally to relen Core toeic fragments. the toxic fragments of ory proteine have 3 domains • Domain I turchione in pore fion chennel formation Domain 2 Involved in receptor recognition Domain 3 benge the receptors. pragnients trings to receptors of the epitheliall wells of the Welle of the wild gut. Mehitrane developes pores @dia result the mehitbras eaming enflex into the walle into the e epithelial. velle. of tons 4420 The The toxic This cause's lyris of death of the insect! epithelial cells leading to other genes of insect resistant- There are several Eurcet resistance genes ushich can also provide - Cry gene gene cannot provide resistant against all type of insects hence some alternateve insecticide genes have been discovered e-CAT-I gene (cow pea trypsin inhibitor 1) Aproteinin gene Lactein gene veal chodectral oxidase gene Vip gene (vegetative insetWidtal protein) VIP CPT-I gene produces proteine that inhibit digestive enzyme - utproteinin game Binhibit serine proteases. Herbicides resistant plants @plaws that wan tokshat herbicides are calleg herbicides resistant plants. peants forth herbies des resistant have been developed b by genetic enginetselly. g-Glyphosate menistant petunia, tomato, Glyphosate tolerate plantes. glyphosate spectrum roungup active herbicedes such as ete. is an glyphal, tumblewee This chemicals enters the the enzyme plant cells and inhibit -Pyruvid shikimate-3.1 5-enol- rynthetase CEPSPA Prynthetase). phosphat Thi seturs only in the chloroplast & synthesized aromatic da from phosphienal pyruvate. (@as glyphosate inhibits EPSP. synthetase aromatic aà cannot be synthesized in plants, and the die loon plant di - Transgeneic plants resistant to several lubricidi I love glyphosate, Glufosinate, selfoniluxcas, have been developed and are commercial cattivat in USA & other countries. phosate resistant plants Aro-geus To produce Glyphosate from agrobacterium, gox-gene fram bacteria & s acxobno- ина Sulfoniturea resistant gene Hrea gene 4 N.tobbiccum are froon Arabidopsis thaliana. (solated of Introduced into a plant to make it The mechaniem follows- glyphosate action es as EPSP+S3P+ Glyphosate EPSPS-SOP-glyphosate complex. ( Anhibition of EPSPS PEP+S3P EPSPL EPSP the Aromah'c Roleg Epips. Luction is as per of Herbicides merish, Me description transgeneic plant given is above figure. Roundup Ready Soyabean: KR soyabean genetically engineered soyabean to which. herbicide Hesistant also known as gene has been introduced. glyphosate tolerate soyabean. Glyphosate kills plant interferring with evential as such as byutheris q evenhal Tyrosine, trptophan. phenylalanine, This ad cannot synthurised oblungot avoid of oblingot of plants 4 by animals and are organisms This an encoded by enzyme (EPSPS). ( Roundup ready soyabeans express a versiong EPSPS syntheare gene from A-tunefacient nohre expresion regulated by 355 promoter pros CMV Thew KR soyabeans beans where sprayed meith glyphosate herbicide, It only kills the weeds and not without affecting H soyabean bean p sean plant. @fint variety of RR sou RR soyabean wehlch was W GST-40-3-2, nus (1994) conimexelally approved en usi Canada (1995) Japan(1996) fs-aprica (2001). there is a new soyabean walled RR-2-yield (RR-24) which claims Increased yield of мор of 7-11% Trenigenic plant welth quality modification) and noble features [crops with Euphroved quality trait] Le pert. eniece ert disease After the development y vesistant and herbicide resistant plants, cortais. plants have been developed with improved quality which wit specific consumers, industries freeds. Such varieties popularly called as designer crops, -This crops possess certain specific features as the requirement. per to meet - This crops quallty have b vitamin tarte etc. specified proteins, content colour, fat, starch Lavor [A] The • first transgenic vaxtety with quality modificat? have been commercial status came mo market as Harr, saur. la variety of It is a variet tomato that shows delayed fruct softening which was approvaved in 1994 In tomects the enzyme polyglacturonase deprogn pectiri uchich ba mayor component of fruct rell walls. This lease to ro fruit and deterioration roftening of fruit. ion fruit quality. quality. 2 Transgenže tomato have been is meintable for food I storage bly 2 approaches ib By technology using antisense RNA tee aming Gene for 1-aminzcyclopropane 1- (ACC) deanlinese wakich degrasjes nec gene to ethylene. transgenics expression g show drastically requced PG & slower frust shortening saur formats has improved flavor f total soluble solids (TSS) in addition to enhance self life the's is possible due to fact alebised & ripen on the that this tomato are alld plant. Hence Janic oreanic acids forest accumulates more forest that in emproves syar theer TSS content. G On ventrest the convertional tomato fruft have to be picked green t hard & treated ubitu ethylene to ripers them. Dantisence ANA technology has been wed to produce a delayed népening of to onats by suppressing ethylens brosynthetis. The antiense contrast o Nec synthase gane es transferred buth tomate which reduces ethylen production oduction less than 17. of normal. Antisense Gene approach:- The term anticente is used to describe inhibition of mRNA transcript translation by hybridicat? of an foktor oligonucleohde to the selected Region of M-RNA. since m-RNA & the serve represent sequence the, oligonucleotide complementary to the M-RNA is called antisense aligonucleotids m any gener the DNA strand oriented as 3-5 In relation to its promotes is transcelbed and the strand is called The mRNA is de chang. ③ The Hem antisense strand. complimentary to that antisense Hemalning DNA 1 of a gene si called sthand of a sense strand which is compechmentary to the antisense trand of a gene. sence stra gene ti :. The boue rez requn MKNA. ene te same that e strand of a gene produced by i't Hence the on-RNA produced by a gere de calley bense RNA An antivenie gene is orientation produced by orientation of reversing orient Enverting ne ene of nelation to ste promoters! the gene RNA produced by this The RNA as the antisense strang gene has same sequence of normal gene knocon as antisenserna. which ei when an antsense gene is present bon same nucleus transcription of the along weith normal endo- genous gene, transcription of 2 genes yoeld anhschse fsense RNA transcripte. The antisente 4 sense RNDA are complementary to each other and undergo pairing to ather double stranded RNA Molecule (RNA-RNA hybrid). [B] Golden nice:- golden lee is a vantety of cexykas sativa (rics) produces through genetic engineering. He a bioportified rice which contain micro- nutrients like vitamin-A to meat the deficiency of chligrens at risk. Drie A deprecency is of beingnon is the single most imp. cause axiong chlegres countries which has per year. in developing affects blac chiegien Ovet A deficiency often cœurs where nice ze the staple food pro-vitid be @Rice endosperm became rice grain B-carotenes. however so does not conten synthesized geranyl- geranyl pyrophosphate which can converted ento vst po vearotene by y enzymete achvity Pk By phytoeneborg syntheare By by phytoens disaturare. > caroten dicaturare. en lycopene cyclase The tran enzyme activity are tranegene providing this enzyme activ transferreg Inb rice transformer. wing Agrobacterium wedst All the transgene are introduced together in single co-bansformation experiment which result transgente rise goldence. popularly called • Golden rice contain good quantity of p-caroten, qohich gives seee grain golgen colour. high as An golden rice B-carotene contain is hi 1.2 dagm/g of endosperm Kinue l on andía 2 tranigene being transferred ento rice vo variety ABT-43 to meet y like swarna, MTU 1010 4 meet vitamin deficieng. en children. A high won high Provitamin A vice variety produced by erasing gorgenrice of the wal transfenk einer of Role of transgende In cological biomemediation:- the gical treatment system to woof biological destroy or warte from a reduce the comm hakadous contaminated refe (terrestrialer @aquabic). the ecofriendly away ollutants to pollutants to cleanup get cleaner 4 greener environment. Blo renudiation Envolves mmlerobes or higher plante uchich are officients to clean the pollutants. gn modern Endustriltered society many un- natural cherideals have been produced and make the environment difficult to leave live. for such cases blorencediation using geneticall engineered netcrobes are used to degrade anch abiatle compoung. C the microbes nekich contain genes transferreg pram different bacterial prasmeds to degrade genebistic xenobidtic called superbry Superbug in a new bacterium ushich contain plasméd and extrachromosomal genetic elements that can with other bacteria to reproduc морир Indenpendently into new ones I share traits with them The fired huperbug ureated to degrage ald was preud was created This superbag by ichakrobarty of co worker? all eating niperbug produced by tramye It is a sil of plasmids ents a single cell of e punte soulof degrade toxic chemicals like octans," xylin, naptheline ete. pušida. @this superbugs are ressed for eleening wilspille. @superbuys are safe for the environnlent cwhich could be an excellent option to deal with polluted, Treatment contaminated rites of different countrie Endustrial efferent of Textiles, zeather, pharmaceutical ete san be made pollutio free. #dible Vaccines: D'Antigens of several pathogene produce immuno- gente response when delever orally such antigene are vaccinert senaidates for edible good for The gene encoding the orally active antigente potein is enolated from the the pathogen f a suitable construct for conihitüteve of kisice specific 2 prepared sehe is ५ The gent construct is introduced ang stably Entegrated into the genomme where it is expr spectes wher antigen to teo ant expressed to produce the The appropriate plant plant containing the antigen may be food raid animal about Immunogaken. or human to br ry-crops used as feed is alfa-alfa.com The edible vaccines are proders plant in uchich orally traget pathogen RA produced from transgenic anhyen & Hie expressed and decunkulated, then it is feed to animal or feed humane for imminóration against pathogen offers' easy deliver Edible vaccine off feeding or lower cost ystems by Reconibinant vaccine.. Another eg of edible vackine is provided by 4-coli heat labile enterotoxin (LT) B subunit (LT-B) expresseq in potato. Heatlabile toxin produces diarrhea & es structurally I funchprally & antigenically very similar to cholera toxin (CT). Both LT FET axe as they excellent oral adjuvent stimulate Emmune response against other antigers feed wwith them. @only feading up 5gm tuber induces fasamune response in mice. Pinunogenecity. [protease & and effectiny fof edible vaccines Industrial enzymes Thh indicates the in Transgene weken introduce anto plante can modify brosynthetic pathways & generates f products. production of value added products like hutrition ridpplements, pharmeccticale pharmecctica wing transgente crops called. de molecular farming The font sonurmercial example brochemical 5 produced is Thrombin inhibitor proten- Hirudin. Tprotease el axe derived from bovins pancrease.. - for the 1st time prodt gene usued a patent for prodexing Higher No teases in transgenic plants. Imo teases dan be expresion of mate obtained using constituent, A seed specific promoter. proteases can be available in the fore foom lyophilized powder ortos steri de candi reth, bottled hotlled for labratory use framer. purposes proteases widel as detergent additives, Ec processing in pharamenical inductiss Eykicare & leather As the bovine slac sccurancs & ter houses are with laughter! 800, prayer of protese mouth diseases, proteases (trypsin) from fer As obtain proteases. plant sources. es- dexived proteins - Tryp zean" Coon dexi TM Lepas is an enzyme that hydrolyzed fate. Other are sub-clan of perform esterase enzyme that digestion, transport, essential roke in diger procening of dietary allside Q Genes encoding espases are present in all kurtain viruses. animals including @most lepases act a specifle position of the ids substrates t glyceras backbone of elpids breakdiscon it into monoglycerides I foa. for exemple Human pancreatic lipase fats in humas digestive breakdovern dietary fats rystem of converts fats ens monoglyceriden Dard fatty acide Upares are cheap 1 emportant catalyst to degrade lipids. Błotech companies has prepared reconebinant lipase enzyme for using baking, detergents Endustries and conve kuel as a and konversion alternative vegetable oile ofle int energy source. Dlused in procession of biodiesel. -Lipases are from animal obtained from a ectacled from and also can be extte sources im microbes. Biosafety :- -Transgenic plants are declared to pare no risk either to human But they yare or to environment. being of conswdered for the elving world as unitafe and - miosafety describe the principle, procedures ang policies to be adapted to ensure the environmental and L and pexionel safety. - cajor reist poserved. The major are- Menereered weedyness. from bantgente plaits flow into di cultivers f Transgene flow related different. > Movixontal transfer ins og organions. ev pamibiesty of wider hou plants. development of viruses range viruses with In verubes resistant f Effect the pesticidal products and non- trargeted brganisme. the mechanism do Poiosafety is defineq developed procedures to ensure the through pakley and procedures environmentally refe of application of biotechnoly which results to develop modified organisms genetically (GMOR). Benefits from transgenic varieties & cultivation of of brangenis Chemicals wise fagro erops reduced the Chemicals the blusechdidis, sprids etc. fungicides Transgente varieties reduces pollution due to use of biodegradle agriculture chemical The crop produced from transgenics is safe. on free from pertteldes relatively prom fre rince it is relativel Herides - Transgentes for improved. Golden reies, flaur-savr nutritional g promote humes health 4 elevate malnutritions -Transgenice yield better crop productions dus to better adeptibility to bione 4 abiotic stren ylet's to sustainable agriculture Pransgenic yield jenic varieties produces Toranigente phardiceuticals, eg- valuable Hiruden, research B-glucomiadare 1 reagents like Aviden 4 B- reagents edible vaccines. It reduce animal se and neinincise pallution. sufferings Disadvantages 6- on human health - effect food prepared from transgenic varieties is referrief as genetically modified food. The potential oisk in product the transgenic are - of transgenic transgenes transgenes or the products may be toxic of ord allerghie products > effect on environnient Tranegente may be a threats to the environment as follows- Mansgenik ① Pransgenk become persistant dece. ctness 4 become fermiers 5_ to their increring of need mind may distributed al theth and damage the ecosystem. - eg: GM heriticides resistant oil seed, malze - and so sugarbest express efther BT-colt toxin of pea-lacks xrop which are or which our highly domesticateg Junable unable to persisting on natural environment without human inter conversion. meect resistant transgente erop may effect non- on target species to part by mod caret transgenic plants kants or organisms may Leve part by non-target effect on them. harmful effect es-CPT-1, chitinase & p-1,3 glicanase, are wed assaying beer where on hondy beer the of these gened chitinare didn't effect it uchile CPT-14 B. 1, 3 glucanese have detrimental effect (sharme 2000) they may encourage theme of chemical they may alter the compochton of khixosphen any phytosphere microspora feord