Ziziphi iiVectors ezifanele zenze ngeGenesis kunye neCloning
Kwi-cloning ye-molecule, i-vector iyimodyuli ye-DNA ekhonza njengomthutha wokudluliselwa okanye ukufakwa kweempawu zomhlaba wangaphandle kwelinye iseli, apho kunokuphinda kuphindwe kunye / okanye kuboniswe. Iimvenge ziphakathi kwezixhobo ezibalulekileyo ze-gene cloning kwaye zona zi luncedo xa zidibanisa uhlobo oluthile lokumakishwa kwejethi olufakela i-molecule ye-bioindicator enokulinganiswa ekuhlolweni kwe-biological ukuqinisekisa ukufakwa kwabo, kunye nokubonakalisa, kwimbutho yokusingatha.
Ngokukodwa, i-cloning vector yi-DNA ethathwe kwintsholongwane, i-plasmid okanye iiseli (zezinto eziphezulu) ukuba zifakwe kunye ne-DNA yangaphandle kwinqununu yeenjongo zokwenza i-cloning. Ekubeni i-vlon cloning ingagcinwa ngokuzinzileyo kwimizimba, i-vector nayo iqulethe izinto ezivumela ukufakelwa okulula okanye ukususwa kwe-DNA. Emva kokufakwa kwi-vlon cloning, isiqwenga se-DNA sinokugqithiselwa kwisinye i-vector esingasetyenziselwa ngakumbi.
Kwezinye iimeko, iintsholongwane zisetyenziselwa ukusulela iibhaktheriya. Ezi zintsholongwane zibizwa ngokuba yi-bacteriophages, okanye i-phage, efutshane. Ama-Retroviruses awona mavecteurs aphezulu kakhulu ekuveliseni izakhi zofuzo kwiiseli zezilwanyana. Iiplasmids, ezizahlulo ze-DNA, zizona zixhobo eziqhelekileyo ezisetyenziselwa ukuzisa iDNA yangaphandle kwiiseli zebhaktheriya. Bahlala bephethe i-antibiotic resistant genes ezingasetyenziselwa ukuvavanya ukubonakalisa i-DNA ye-plasmid, kwiipiritiki ze-petri.
I-Gene idluliselwa kwiiseli zezityalo ngokuqhelekileyo eyenziwa ngokusebenzisa i-bacterium yomhlaba i- Agrobacterium tumefaciens , eyenziwa njengemvolisi kwaye ifake iplasmid enkulu kwiseli yomkhosi. Kuphela kuphela iiseli eziqulethe i-cloning vector ziyakhula xa i-antibiotics ikhona.
IiNdidi eziMkhulu zoVimba beCloning
Iintlobo ezi-6 eziphambili ze-vectors zi:
- Plasmid. I-DNA echitshiyorosomal DNA eyazimela ngokuzimela ngaphakathi kweseli yebhakteria. Iiplasmids ngokubanzi zinombhalo ophezulu wekopopi, njenge-pUC19 enekopopi yenani lama-500-700 ikopi nganye kwiseli.
- Phage. Ama-molecule e-DNA ahlukeneyo aphuma kwi-bacteriophage lambda. Unokutshintshwa kwi-DNA yangaphandle ngaphandle kokuphazamisa umjikelezo wobomi bawo.
- Cosmids. Enye i-molecule ye-DNA i-extrachromosomal i-DNA edibanisa iimpawu ze-plasmids kunye ne-phage.
- Ama-Chromosomes aseBactter Artificial Artificial. Ngokusekelwe kwi-mini-F plasmids ye-bacteria.
- I-Chromosomes Artificial Artificial. Le chromosome engumgubo equkethe i-telomeres (iilaphu ezilahlayo ekupheleni kwee-kromosomes ezicatshulwa ngexesha lokuhlukana kweeseli) ngemvelaphi yokuphindaphinda, i-leaven centromere (inxalenye ye-chromosome edibanisa i-chromatids nodade okanye i-dyad), kunye nomakethi okhethiweyo ukuchongwa kwimvubelo yamaseli.
I-Chromosome Yomhlaba Yomntu. Olu hlobo lwevolisi inokubaluleka ekunikezeni i-gene kwiiseli zomntu, kunye nesixhobo sokuhlola iinjongo kunye nokwenza umsebenzi we-chromosome yomntu. Iyakwazi ukuthwala isahluko esikhulu seDNA.
Zonke izitshixo ezibunjiweyo zinomvelaphi wokuziphendulela (i-replicator), indawo ye-cloning (efumaneka apho ukufakwa kwe-DNA yangaphandle akuphazamisi ukuphindaphinda okanye ukungasebenzi kweempawu zokubaluleka), kunye nomakethi okhethiweyo (ngokuqhelekileyo i-gene ehlinzekela ukulwa nokulwa ne-antibiotics.)