Maplasmid muMolecular Genetics
Maplasmid ndeimwe yezvinhu zvakakosha zvemajini mumajini emazuva ano emolecular. Pasinei nehukuru hwawo hudiki kana tichienzanisa nemachromosome, maplasmid anoita basa guru mukunzwisisa mashandiro emajini, kushandisa DNA, uye kutogadzira zvigadzirwa zvebiotechnology zvakaita se recombinant insulin. Munyaya ye microbiology, maplasmid anowanzo kuwanikwa mumabhakitiriya nezvimwe zvipenyu zvakawanda se extrachromosome DNA inokwanisa kuzvinyora. Kuvapo kwemaplasmid hakusi kungo "wedzera" kune majini; anowanzo sarudza hunhu hwakakosha senge kuramba maantibiotic uye kugona kugadzira chepfu kana ma enzymes.
Tsanangudzo uye hunhu hweplasmids
Kazhinji, plasmid imorekuru yeDNA yakatenderera, ine tambo mbiri yakaparadzana neDNA huru yechromosome. Maplasmid akasiyana muhukuru, kubva pazviuru zvishoma zvema base pairs (kilobases) kusvika kumazana ema kilobases, zvichienderana nerudzi. Nekuti anogona kuzvinyora ega, maplasmid ane nzira yakakosha inonzi orig of replication (ori). Izvi zvinobvumira plasmid kubereka mukati memusero wehost pasina kuvimba zvachose ne chromosome replication.
Huwandu hwemakopi eplasmid musero rimwe chete hunosiyanawo. Mamwe maplasmid ane makopi akawanda (semuenzaniso, makumi kusvika kumazana emakopi pasero rega rega) uye mamwe ane makopi mashoma (makopi mashoma chete). Musiyano uyu wakakosha mu molecular genetics nekuti unokanganisa mwero wekutaura kwemajini anotakurwa neplasmid uye kugadzikana kwawo panguva yekupatsanurwa kwesero.
Mhando dzeplasmids zvichienderana nebasa radzo
Maplasmid anogona kuiswa mumapoka zvichienderana nemajini aari pamwe nekubatsira kwawo pakurarama kana kuti kukunda mabhakitiriya.
1. Kudzivirira plasmid (R plasmid)
Maplasmid aya ane majini anoita kuti mabhakitiriya asarambe mishonga inorwisa mabhakitiriya. Muenzaniso ijini rinogadzira beta-lactamase enzyme, iyo inogona kupwanya mishonga inorwisa mabhakitiriya yebeta-lactam yakaita sepenicillin. Maplasmid eR ane basa guru munyaya dzezvekurapa nekuti anogona kupararira pakati pemabhakitiriya uye kukurumidzisa kubuda kwemabhakitiriya asingarambe mishonga.
2. Plasmidhi yekubereka (F plasmid)
Plasmidhi yeF inoita basa guru mukubatana kweDNA, kutamiswa kweDNA pakati pemabhakitiriya kuburikidza nekubatana zvakananga. Mabhakitiriya ane F plasmid anokwanisa kugadzira maumbirwo akafanana nepili (bonde pili) kuti abatanidzwe kune mamwe mabhakitiriya uye kutamisa maplasmid awo. Kubatana ndeimwe yenzira huru dzekupararira kwemajini, kusanganisira majini ekudzivirira.
3. Maplasmid ane utachiona
Maplasmid aya ane majini anowedzera kugona kwebhakitiriya kukonzera zvirwere, zvakaita semajini etoxin, zvinhu zvinonamatira, kana mapuroteni anobatsira kudzivirira immune system yemuviri wemunhu. Saka, maplasmid ane hutachiona chinhu chakakosha mukukonzera mamwe mabhakitiriya ane hutachiona.
4. Maplasmid emuviri (anoparadza)
Maplasmid aya anogonesa mabhakitiriya kugadzira zvinhu zvisingawanzoitiki, zvakaita semahydrocarbon kana mishonga yekuuraya zvipembenene. Mu biotechnology yezvakatipoteredza, maplasmid emetabolism ndiwo hwaro hwekushandisa mabhakitiriya mukugadzirisa zvinhu zvipenyu.
Basa replasmids mu molecular genetics
Mukuongorora majini emamorekuru, maplasmid haasi zvinhu zvekudzidza chete asiwo zvishandiso zvikuru. Maplasmid anoshanda semavector ekupinza majini mumasero uye kudzora kuratidzwa kwawo. Tsvagiridzo mumaitiro ekudzokororwa, kutonga majini, uye kutamiswa kwemajini kwakatwasuka kunoenderana zvakanyanya nekushandiswa kwemaplasmid senzira dzemuenzaniso.
Maplasmid anotsigirawo zvakanyanya nzira dzekutanga dzakadai sekugadzira magene cloning. Nekuisa chidimbu cheDNA chakanangana nacho muplasmid, vaongorori vanogona kuwedzera DNA iyoyo zvakanyanya nekungokura mabhakitiriya anotakura. Uyezve, maplasmid anogona kugadzirwa kuti agadzire mapuroteni (express) kubva kumagene akanangana nawo murabhoritari kana muindasitiri.
Zvikamu zvakakosha zvevector plasmids
Maplasmid anoshandiswa mukugadzira majini anowanzo kuve maplasmid akagadziriswa kuti ave nyore kushandisa semavector. Zvikamu zvikuru zvevector plasmid zvinosanganisira:
- Kwakatangira kudzokororwa (ori): kunoita kuti plasmid ikwanise kudzokororwa muhosi.
– Chiratidzo chinosarudzwa: jini rechiratidzo chekusarudza, kazhinji jini risingakwanise kushandiswa nemishonga inorwisa mabhakitiriya (semuenzaniso ampicillin kana kanamycin) kuitira kuti mabhakitiriya ane plasmid chete ndiwo anogona kukura pamichina yakasarudzwa.
– Nzvimbo yekuunganidza magene akawanda (MCS): nzvimbo ine nzvimbo dzakawanda dzekudzivirira kuti DNA ishandiswe uchishandisa maenzyme ekudzivirira.
– Zvinosimudzira uye zvinhu zvinodzora: zvinodiwa kana maplasmid achishandiswa pakuratidza majini. Zvinosimudzira zvine simba (senge T7 kana lac promoter) zvinogona kukurudzira kugadzirwa kwemapuroteni akawanda.
– Gene remutori wenhau (sarudzo): gene remutori wenhau rakadai seGFP kana lacZ kuti zviite kuti zvikwanise kuzivikanwa kwemapoka anotakura DNA zvinobudirira.
Kugadzirwa kweplasmid kwakakodzera ndiko kunosarudza kubudirira kwechiedzo, zvese pakugadzira mapuroteni ari nyore uye pakugadzirwa kwemapuroteni akawanda.
Maplasmids ari mukuita cloning uye DNA recombination
Tekinoroji yeDNA recombinant inoshandisa maplasmid senzira dzinotakura majini. Maitiro aya anowanzo sanganisira kuparadzanisa plasmid, kucheka plasmid neDNA retarget ne restriction enzymes, kubatanidza zvidimbu uchishandisa DNA ligase, uye wozoisa recombinant plasmid mumabhakitiriya kuburikidza nekushandura. Makoroni emabhakitiriya anotakura plasmid zvinobudirira anogona kusarudzwa uchishandisa zviratidzo zvekusarudza.
Chinhu chikuru chinobatsira maplasmid ndechekuti zviri nyore kugadzira zvakawanda uye kugadzikana kwawo mukati memabhakitiriya. Mukati mehusiku humwe chete, bhakitiriya rinogona kugadzira maplasmid akakwana ekuongorora kwakasiyana-siyana, akadai sePCR, sequencing, kana transfection mumasero eukaryotic.
Mashandisirwo eplasmids mu biotechnology nemushonga
Maplasmid anoshandiswa zvakanyanya. Muindasitiri yemishonga, maplasmid anobatsira mukugadzirwa kwemapuroteni ekurapa, akadai se insulin yemunhu, mahormone ekukura, uye ma antibodies e recombinant. Mukutsvagurudza kwekutanga, maplasmid anoshandiswa kudzidza mashandiro emajini kuburikidza nekuwedzera kwehuwandu hwemasero kana mutagenesis inotungamirirwa nenzvimbo.
Maplasmid anoshandiswawo mukugadzira majekiseni eDNA, nzira yekudzivirira inoshandisa maplasmid ane majini eantigen. Kana maplasmid akapinda mumasero emuviri, anogadzira maantigen uye anokonzera mhinduro yekudzivirira muviri. Kunyange zvazvo kugadzirwa kwejekiseni reDNA kuchitarisana nematambudziko ehunyanzvi uye emutemo, pfungwa iyi inoratidza kuchinjika kwemaplasmid semapuratifomu ejini.
Mukurapa majini, dzimwe nguva maplasmid anoshandiswa senzira isina utachiona yekutakura majini ekurapa. Zvakanakira zvawo ndezvekuti akachengeteka pane mavhairasi mune zvimwe zvinhu, kunyangwe kupinda zvakanaka mumasero anotarisirwa uye kugadzirwa kwemajini kuchiramba kuri dambudziko riri kuenderera mberi nekutsvaga.
Matambudziko etsika nematambudziko
Kushandiswa kwemaplasmid, kunyanya ayo ane majini ekudzivirira mabhakitiriya sezviratidzo, kunokonzera kushushikana nezvekupararira kwekudzivirira mabhakitiriya kana maplasmid akaburitswa munzvimbo yakatenderedza kana kutamisirwa kune mamwe mabhakitiriya. Nokudaro, vaongorori neindasitiri ye biotechnology vanofanirwa kushandisa nzira dzakasimba dzekuchengetedza mabhakitiriya. Dzimwe nzira dzekusarudza dzisingashandisi maantibiotic dziri kugadzirwa parizvino, dzakadai se metabolic-based markers kana positive-negative selection systems.
Kubva pamaonero etsika, hunyanzvi hwekugadzira majini huchishandisa maplasmid hunoda mutoro mukushandiswa nekuparadzirwa kwezvisikwa zvakagadziriswa majini. Mitemo yakasimba inodiwa kuti kuve nechokwadi chekuti hunyanzvi hwebiotechnology hunobatsira pasina kukonzera njodzi dzisingadzoreki kuhutano nenharaunda.
Mhedziso
Maplasmid zvinhu zveDNA zviri kunze kwemachromosome zvinoita basa guru mukugadzirwa kwemajini emamorekuru. Zvese zviri zviviri zvinhu zvechisikigo zvinopesvedzera kugadziriswa kwemabhakitiriya uye semavectors ekugadzira majini, maplasmid ndiwo hwaro hwenzira dzakasiyana dzemazuva ano dzakadai sekugadzira macroning, protein expression, uye kugadzirwa kwema biomolecules ekurapa. Nekunzwisisa kwakadzama kwemaumbirwo eplasmid, mashandiro, uye nzira, vaongorori vanogona kuramba vachigadzira mashandisirwo akachengeteka, anoshanda, uye anobatsira ebiotechnology kuvanhu.
