Dab tsi yog txoj kev purification ntawm lubprotein purification system? Nws yog ib qho tsim nyog yuav tsum paub txog cov txheej txheem DNA ntawm cov protein ua kom huv, kom pom cov hlwb lossis cov ntaub so ntswg uas tau nthuav tawm ntau dhau ntawm cov noob caj noob ces, thiab tsim cov gene primers kom nthuav dav arf ntawm lub hom phiaj DNA tawg. Qhov no yog qhov hu ua kev nrhiav tau ntawm lub hom phiaj gene fragments.

Kev tsim kho ntawm kev qhia vector: Hauv prokaryotic lossis eukaryotic qhia vector nrog cov yam ntxwv ntawm cov noob tau qhia nws tus kheej, qhov teeb meem tseem ceeb ntawm cov kauj ruam no yog tsim cov plasmid thiab cov noob paj, thiab cov kab lus qhia. Lub sijhawm qhia prokaryotic yog luv luv, tus nqi qis, thiab ntau qhov kev qhia yog qhov tseem ceeb; cov noob tsis tau qhia hauv E. coli, thiab qhov teeb meem tshwm sim hauv codon optimization. Hauv kev xav txog kev ua haujlwm zoo dua thiab ua kom huv ntawm cov protein, cov kws tshawb fawb tau xaiv los qhia hauv Pichi poov xab. Kev ua tiav ntawm codon optimization yog qhov tseem ceeb.

Dab tsi yog txoj kev purification ntawm cov protein purification system:

1. Dej nag.

2. Electrophoresis: Ib qho protein uas tau them yog siab dua lossis qis dua nws qhov isoelectric point thiab tuaj yeem txav mus rau qhov tsis zoo electrode lossis qhov zoo electrode ntawm cov hluav taws xob hauv hluav taws xob. Txhawb zaj duab xis electrophoresis, electrophoresis, thiab lwm yam.

3. Dialysis: Ib txoj hauv kev uas siv ob lub hnab lim ntshav los cais cov molecules loj ntawm cov proteins thiab cov organic molecular me me.

4. Chromatography: Ion pauv chromatography siv cov khoom pub dawb ntawm cov protein. Raws li qhov tshwj xeeb pH, cov nqi thiab cov khoom ntawm cov protein sib txawv, thiab lawv tuaj yeem sib cais los ntawm ion exchange chromatography. Hauv anion pauv chromatography, cov proteins uas tsis muaj zog tsis zoo yog eluted ua ntej. Molecular sieves, tseem hu ua gel filtration. Cov protein me me nkag mus rau hauv qhov hws thiab nyob hauv lawv ntev. Cov protein ntau tsis tuaj yeem nkag mus rau hauv lub qhov hws thiab ntws tawm ncaj qha.

5. Dab tsi yog txoj kev purification ntawm lubprotein purification system? Ultracentrifugation: Protein purification tuaj yeem siv los txiav txim siab qhov hnyav molecular thiab tuaj yeem siv los ua cov protein. Kev tsim cov proteins uas muaj qhov sib txawv yog sib cais.