Li-plasmid ho Liphatsa tsa Lefutso tsa Limolek'hule
Li-plasmid ke e 'ngoe ea likarolo tsa bohlokoa ka ho fetisisa tsa lefutso liphatseng tsa lefutso tsa sejoale-joale tsa limolek'hule. Ho sa tsotellehe boholo ba tsona bo bonyenyane ha li bapisoa le li-chromosome, li-plasmid li bapala karolo ea bohlokoa ho utloisiseng ts'ebetso ea liphatsa tsa lefutso, ho laola DNA, esita le ho hlahisa lihlahisoa tsa biotechnology tse kang insulin e kopantsoeng. Moelelong oa microbiology, li-plasmid hangata li fumanoa libaktheria le linthong tse ling tse 'maloa tse phelang e le extrachromosome DNA e khonang ho ikatisa. Boteng ba li-plasmid ha se feela "tlatsetso" ho thepa ea liphatsa tsa lefutso; hangata li khetholla litšobotsi tsa bohlokoa tse kang ho hanyetsa lithibela-mafu le bokhoni ba ho hlahisa chefo e itseng kapa li-enzyme.
Tlhaloso le litšobotsi tsa li-plasmid
Ka kakaretso, plasmid ke molek'hule ea DNA e chitja, e nang le mela e 'meli e arohaneng le DNA e kholo ea chromosome. Li-plasmid li fapana ka boholo, ho tloha ho lipara tse likete tse 'maloa tsa motheo (li-kilobase) ho isa ho li-kilobase tse makholo, ho latela mofuta oa tsona. Hobane li ka ikatisa ka botsona, li-plasmid li na le tatellano e khethehileng e bitsoang tšimoloho ea ho ikatisa (ori). Sena se lumella plasmid ho ikatisa ka har'a sele ea moamoheli ntle le ho itšetleha ka botlalo ka ho ikatisa ha chromosome.
Palo ea likopi tsa plasmid ka seleng e le 'ngoe le eona ea fapana. Li-plasmid tse ling li na le lipalo tse phahameng tsa likopi (mohlala, likopi tse mashome ho isa ho tse makholo ka sele) 'me tse ling li na le lipalo tse tlase tsa likopi (likopi tse 'maloa feela). Phapang ena e bohlokoa ho liphatsa tsa lefutso tsa limolek'hule hobane e ama boemo ba tlhahiso ea liphatsa tsa lefutso tse jereng plasmid le botsitso ba tsona nakong ea karohano ea sele.
Mefuta ea li-plasmid ho latela mosebetsi oa tsona
Li-plasmid li ka hlophisoa ho latela liphatsa tsa lefutso tseo li nang le tsona le monehelo oa tsona ho pholoheng kapa ho phahameleng baktheria.
1. Plasmidi e hanyetsanang (Plasmidi e hanyetsanang)
Li-plasmid tsena li na le liphatsa tsa lefutso tse fanang ka khanyetso ea libaktheria ho lithibela-mafu tse itseng. Mohlala ke jini e kenyang enzyme ea beta-lactamase, e ka senyang lithibela-mafu tsa beta-lactam joalo ka penicillin. Li-plasmid tsa R li tšoenyehile haholo lefatšeng la bongaka hobane li ka ata lipakeng tsa libaktheria le ho potlakisa ho hlaha ha libaktheria tse hanyetsanang le mefuta e mengata.
2. Plasmidi ea monono (Plasmidi ea F)
Plasmidi ya F e bapala karolo ho kopanyeng, phetiso ya DNA pakeng tsa dibaktheria ka ho kopana ka ho toba. Dibaktheria tse nang le plasmid ya F di kgona ho etsa dibopeho tse kang pili (pili ya bong) ho kgomarela dibaktheria tse ding le ho fetisetsa diplasmid tsa tsona. Ho kopanyeng ke e nngwe ya mekgwa ya mantlha ya ho ata ha dijini, ho kenyeletswa le dijini tse hanyetsanang.
3. Li-plasmid tse bakang kokoana-hloko
Li-plasmid tsena li na le liphatsa tsa lefutso tse ntlafatsang bokhoni ba baktheria ba ho baka mafu, joalo ka liphatsa tsa lefutso tsa chefo, lintlha tsa ho khomarela, kapa liprotheine tse thusang ho qoba sesole sa 'mele sa moamoheli. Kahoo, li-plasmid tse kotsi ke karolo ea bohlokoa ho pathogenesis ea libaktheria tse ling tse bakang mafu.
4. Li-plasmid tsa metabolism (tse senyang)
Li-plasmid tsena li nolofalletsa libaktheria ho fetola metsoako e sa tloaelehang, joalo ka li-hydrocarbon kapa chefo e bolaeang likokoanyana. Theknolojing ea baeloji ea tikoloho, li-plasmid tsa metabolism ke motheo oa ho sebelisa libaktheria bakeng sa ho lokisa likokoana-hloko.
Karolo ea li-plasmid liphatseng tsa lefutso tsa limolek'hule
Thutong ea liphatsa tsa lefutso tsa limolek'hule, li-plasmid ha se lintho tse ithutoang feela empa hape ke lisebelisoa tsa bohlokoa. Li-plasmid li sebetsa e le li-vector bakeng sa ho kenya liphatsa tsa lefutso liseleng le ho laola tlhahiso ea tsona. Patlisiso mabapi le mekhoa ea ho ikatisa, taolo ea liphatsa tsa lefutso, le phetisetso ea liphatsa tsa lefutso tse rapameng e itšetlehile haholo ka tšebeliso ea li-plasmid e le litsamaiso tsa mohlala.
Li-plasmid li boetse li tšehetsa mekhoa ea motheo e kang ho kopanya liphatsa tsa lefutso. Ka ho kenya karolo ea DNA e reretsoeng ho e kenya ka har'a plasmid, bafuputsi ba ka holisa DNA eo ka bongata bo boholo ka ho holisa baktheria e jereng. Ho feta moo, li-plasmid li ka etsoa ho hlahisa (ho hlahisa) liprotheine ho tsoa liphatseng tsa lefutso tse reretsoeng laboratoring kapa sekala sa indasteri.
Likarolo tsa bohlokoa tsa li-plasmid tsa vector
Li-plasmid tse sebelisoang boenjiniereng ba liphatsa tsa lefutso hangata ke li-plasmid tse fetotsoeng hore li sebelisoe habonolo e le li-vector. Likarolo tse ka sehloohong tsa vector plasmid li kenyelletsa:
– Tšimoloho ea ho pheta-pheta (ori): e etsa bonnete ba hore plasmid e ka ipheta ka har'a moamoheli.
– Letshwao le ka kgethwang: jini ya letshwao la kgetho, hangata jini ya ho hanyetsa dithibela-mafu (mohlala, ampicillin kapa kanamycin) e le hore ke dibaktheria tse jereng plasmid feela tse ka holang mefuteng e kgethilweng.
– Sebaka sa ho kopanya di-clone tse ngata (MCS): sebaka se nang le dibaka tse ngata tsa thibelo ho nolofatsa ho kenngwa ha DNA e shebilweng ho sebediswa dienzyme tsa thibelo.
– Li-promoter le likarolo tsa taolo: lia hlokahala ha li-plasmid li sebelisoa bakeng sa tlhahiso ea liphatsa tsa lefutso. Li-promoter tse matla (tse kang T7 kapa lac promoter) li ka khanna tlhahiso e phahameng ea protheine.
– Gene ea moqolotsi oa litaba (ha ho hlokahale): gene ea moqolotsi oa litaba joalo ka GFP kapa lacZ ho nolofatsa ho tsebahatsa likolone tse jarang DNA ka katleho.
Moralo o nepahetseng oa plasmid o etsa qeto ea katleho ea teko, ka bobeli bakeng sa ho etsa kloning e bonolo le tlhahiso ea protheine e kholo.
Li-plasmids tse ntseng li etsoa ka mokhoa oa ho kopanya le ho kopanya DNA
Theknoloji ea DNA e kopantsoeng e sebelisa li-plasmid e le likoloi tse tsamaisang liphatsa tsa lefutso. Ts'ebetso ena ka kakaretso e kenyelletsa ho arola plasmid, ho khaola plasmid le DNA e reretsoeng ka li-enzyme tse thibelang, ho kopanya likotoana ho sebelisoa li-ligase tsa DNA, ebe ho kenngoa plasmid e kopantsoeng ka har'a libaktheria ka phetoho. Likolone tsa baktheria tse jarang plasmid ka katleho li ka khethoa ho sebelisoa matšoao a khetho.
Molemo o moholo oa li-plasmid ke bonolo ba tsona ba ho hlahisa ka bongata le botsitso ba tsona ka har'a libaktheria. Ka bosiu bo le bong feela, setso sa baktheria se ka hlahisa li-plasmid tse lekaneng bakeng sa litlhahlobo tse fapaneng, joalo ka PCR, tatellano, kapa phetisetso liseleng tsa eukaryotic.
Tšebeliso ea li-plasmid ho theknoloji ea baeloji le bongaka
Li-plasmid li na le ts'ebeliso e pharaletseng. Indastering ea meriana, li-plasmid li thusa tlhahisong ea liprotheine tsa kalafo, joalo ka insulin ea motho, lihormone tsa kholo, le li-antibodies tse kopantsoeng hape. Phuputsong ea motheo, li-plasmid li sebelisoa ho ithuta ts'ebetso ea liphatsa tsa lefutso ka ho hlahisa ho feteletseng kapa phetoho e lebisitsoeng sebakeng.
Li-plasmid li boetse li sebelisoa ho nts'etsopele ea liente tsa DNA, e leng mokhoa oa ho enta o sebelisang li-plasmid tse jereng liphatsa tsa lefutso tsa antigen. Ha li-plasmid li kena liseleng tsa 'mele, li hlahisa li-antigen, e leng se bakang karabelo ea' mele ea ho itšireletsa mafung. Leha nts'etsopele ea liente tsa DNA e tobane le liphephetso tsa tekheniki le tsa taolo, khopolo ena e bontša ho tenyetseha ha li-plasmid e le liforomo tsa liphatsa tsa lefutso.
Kalafong ea liphatsa tsa lefutso, li-plasmid ka linako tse ling li sebelisoa e le mokhoa o mong o seng oa vaerase bakeng sa ho jara liphatsa tsa lefutso tsa phekolo. Molemo oa tsona ke hore li sireletsehile ho feta li-vector tsa vaerase likarolong tse itseng, leha ho le joalo ho kena ka katleho liseleng tse reretsoeng le tlhahiso ea liphatsa tsa lefutso e ntse e le liphephetso tse ntseng li tsoela pele lipatlisisong.
Liphephetso le mathata a boitšoaro
Tšebeliso ea li-plasmid, haholo-holo tse nang le liphatsa tsa lefutso tsa ho hanyetsa lithibela-mafu e le matšoao, e baka matšoenyeho mabapi le ho ata ha ho hanyetsa haeba li-plasmid li lokolloa tikolohong kapa li fetisetsoa libaktheria tse ling. Ka hona, bafuputsi le indasteri ea theknoloji ea baeloji ba hloka ho kenya tšebetsong mekhoa e thata ea polokeho ea likokoana-hloko. Mekhoa e meng ea khetho e sa itšetleheng ka lithibela-mafu e ntse e ntlafatsoa hona joale, joalo ka matšoao a thehiloeng metabolism kapa mekhoa ea khetho e ntle le e mpe.
Ho ya ka pono ya boitshwaro, boenjiniere ba lefutso bo sebedisang di-plasmid bo hloka boikarabelo tshebedisong le kabong ya diphedi tse fetotsweng ka lefutso. Ho hlokahala melao e thata ho netefatsa hore boqapi ba biotechnology bo fana ka melemo ntle le ho baka dikotsi tse sa laoleheng bophelong bo botle le tikolohong.
Qetello
Li-plasmid ke likarolo tsa DNA tse kantle ho chromosome tse bapalang karolo ea bohlokoa liphatsa tsa lefutso tsa limolek'hule. Ka bobeli e le mabaka a tlhaho a susumetsang ho ikamahanya le libaktheria le e le li-vector bakeng sa boenjiniere ba liphatsa tsa lefutso, li-plasmid ke motheo oa mekhoa e fapaneng ea sejoale-joale e kang ho kopanya, ho hlahisa protheine le tlhahiso ea li-biomolecule tsa kalafo. Ka kutloisiso e tebileng ea sebopeho sa plasmid, mosebetsi le mekhoa, bafuputsi ba ka tsoela pele ho nts'etsapele lits'ebetso tse sireletsehileng, tse sebetsang hantle le tse molemo haholoanyane tsa theknoloji ea baeloji bakeng sa batho.
