Mokhoa oa tlhoekiso eatsamaiso ea ho hloekisa protheine? Hoa hlokahala ho tseba tatellano ea li-coding ea DNA ea protheine e hloekisitsoeng, ho bona hore na ke lisele life kapa lisele tse hlahisoang ka mokhoa o fetelletseng mofuteng oo ho lebelloang, le ho rala li-primer tsa lefutso ho holisa arf ea sekhechana sa DNA. Sena ke seo ho thoeng ke ho fumana likaroloana tsa liphatsa tsa lefutso.

Kaho ea vector ea polelo: Ka har'a prokaryotic kapa eukaryotic expression vector e nang le litšobotsi tsa mofuta oa mofuta o fumanoeng, bothata bo boholo ba mohato ona ke ho theha plasmid le mofuta oa thahasello, le sistimi ea polelo. Nako ea polelo ea prokaryotic e khuts'oane, litšenyehelo li tlase, 'me palo e kholo ea polelo ke ntho e tlang pele; jini ha e hlalosoe ho E. coli, 'me bothata bo etsahala ka ho ntlafatsa codon. Ha ho nahanoa ka ts'ebetso e ntle le tlhoekiso ea protheine, bafuputsi ba ile ba khetha ho hlahisa tomoso ea Pichi. Polelo e atlehileng ea ntlafatso ea codon e bohlokoa.

Mokhoa oa ho hloekisa protheine ke ofe:

1. Pula.

2. Electrophoresis: Protheine e qosoang e phahame kapa e tlaase ho feta ntlha ea eona ea isoelectric 'me e ka isoa ho electrode e mpe kapa electrode e ntle ea tšimo ea motlakase sebakeng sa motlakase. Tšehetsa filimi ea electrophoresis, electrophoresis, joalo-joalo.

3. Dialysis: Mokhoa o sebelisang mekotla e 'meli ea dialysis ho arola limolek'hule tse kholo ho liprotheine le metsoako e nyenyane ea limolek'hule.

4. Chromatography: Ion exchange chromatography e sebelisa thepa ea mahala ea liprotheine. Tlas'a pH e itseng, litefiso le thepa ea liprotheine li fapane, 'me li ka aroloa ka chromatography ea ion exchange. Ho anion exchange chromatography, liprotheine tse nang le matla a fokolang a fokolang li hlakoloa pele. Li-sieve tsa limolek'hule, tse tsejoang hape e le filtration ea gel. Liprotheine tse nyenyane li kena ka har'a li-pores 'me li lula ho tsona nako e telele. Liprotheine tse kholo ha li khone ho kena ka li-pores ebe li phalla ka ho toba.

5. Mokhoa oa tlhoekiso eatsamaiso ea ho hloekisa protheine? Ultracentrifugation: Ho hloekisa protheine ho ka sebelisoa ho fumana boima ba limolek'hule mme e ka sebelisoa e le protheine. Sebopeho sa liprotheine tse nang le li-density tse fapaneng li arohane.