BiorecHnovogy Pancipres Tech. baset on” biolgy PROCESSES pihen used th a91i 5 food sclence fea $ 5: live orb anise Biotechnology deale with techniques using“ 'o, fi TON oR Intepration of notural _scie =i eet mmolecula |ptoducts! x \setvfces - enzymes ovpanism je produce products a ofall presses to humans ne % oxpanisir ek or y analogues \ Bor ecTNoLugy As PER glotech AMS Sa *s Dey PROCESSES cre fo Such pigceatt ee call modified vp to . Modern = achieve S$ me ove "set gia pea ale ew ; . 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[ ennyrnesy CRecombina ate one} / trimeric DNA as propertl fom. waudtinic SOUTCES(oncePTUAL DEVAOPMENT OF PRINCIPLES of GENETIC ENGINEERING in plant «Traditional hybridisation proced utes uge d g, anima | breeding lead tp incluslor & multiplication © undeslifble oe aon waif est t able Qe Z which include 5 Techniques of genetic enpineevin 6 treation °F nate recombinant DNA suse of gene cloning Q gem tran fo ery xhis vimita tf om: Ov econ1.3 What is gene cloning? What exactly is gene cloning? The easiest way to answer this question is to follow through the steps in a gene cloning experiment (Figure 1.1): 1 A fragment of DNA, containing the gene to be cloned, is inserted into a circular DNA molecule called a vector, to produce a recombinant DNA molecule. The vector transports the gene into a host cell, which is usually a bacterium, although other types of living cell can be used. Within the host cell the vector multiplies, producing numerous identical copies, not only of itself but also of the gene that it carries. When the host cell divides, copies of the recombinant DNA molecule are passed to the progeny and further vector replication takes place. After a large number of cell divisions, a colony, or clone, of identical host cells is produced. Each cell in the clone contains one or more copies of the recombinant DNA molecule; the gene carried by the recombinant molecule is now said to be cloned.1 Construction of a recombinant DNA molecule Figure 1.1 ‘The basic steps in gene cloning. Bacterial colonies rowing on solid mediimThere ave 3 baci me 4 1 ey nae O Tdentificati on of BNA with dest rable. gene &) Entiodudion of identified DNA into host 6) Maintenance of iptieducecL DNA in host Z transfer of DNA to ir prope, Teors OF RELONB BAA “TECHNOWeY ; y Restriction enzyme e enwyme iv) Potyrner envyme 9) ligase enzyme (D DNA marnpuleti¥ 4) Vectors GB) Host organism - pan Pu inte ENZYME \ q) Pe nee a a Recirictia™ enzyme@ Res raictioN Guzynes : (RE? false Known as Moleculay scissov (ge , tn Escherichia eli pec tvicti vg e view {sola teal tc scalpel | chemical Knives: Discovery: in 1463 Cr enzyme ‘responsible fot pores of bacteti ophag Gom Elli. ' wet —- as” one of iF on of it” oo 7 L cut DNA - Monirication’ added CNA of bacteriophage) methyl qoup {named + preverrt -rect- Gray ResrricTioN ENDONUCLEASE endo nucleare. por cleaving €:Col! pya: B CPaeventor. of patietioplone | muttipliati orb in bacteria) — Phage Inj ects DNA Inte bactevium —_ Rechictio” ane endonuceste cleave \ tnatttetd acetal DNA =e mot cleaved Me Me | ont oid pe aa” | 40 42008 w= He Me we \ - asim Reog - SeQy- methylated :(a) Restriction of phage DNA Phage injects DNA into a bacterium Restriction Phage DNA endonucleases bind is cleaved to the phage DNA and inactivated (b) Bacterial DNA is not cleaved Bacterial DNA Me Me Restriction endonucleases cannot bind to the recognition N Me sequence Me Recognition sequences are methylated Figure 4.8 The function of a restriction endonuclease in a bacterial cell: (a) phage DNA is cleaved, but (b) bacterial DNA is not.Features: Restriction enzyme are obtained only fron prokaryotes a ; lt ie thety natural defense mechanism agai bact eviophoge infection. - Tk seve as chemical Knive Into defined fragment fo cut genes CDNA) ) > £2) RecrercTioN ENDONUCLEASE pomed 24 oe ‘\ vestricti capability of cut pability © 4 growths “— viral genome 60 NA) attacking backenia interally into several ieces . @-Re belong > nuclease C Hydsolace’ - Sey 4($) Cut both strands of DB © oe tee phate backhoe) (sugar phos He Re pvr void bY “4 ognisin partie plat specific seqeente cence po t ty pose: er ess Qoo vestriction © 7 230 Stains. 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