發(fā)布時(shí)間：2018-04-17 10:21

  本文選題：奶牛 + NF-κB��； 參考：《東北農(nóng)業(yè)大學(xué)學(xué)報(bào)》2016年09期

【摘要】：NF-κB1是一類重要核轉(zhuǎn)錄因子,參與多種基因表達(dá)調(diào)控。NF-κB1能與奶牛乳腺中乳合成調(diào)控相關(guān)基因結(jié)合,發(fā)揮泌乳調(diào)節(jié)作用。研究應(yīng)用PCR方法擴(kuò)增奶牛NF-κB1基因編碼序列,通過(guò)Eco RⅠ和XhoⅠ酶切位點(diǎn)將其定向插入p GEX-4T-1載體中,構(gòu)建原核表達(dá)重組質(zhì)粒p GEX-4T-1-NF-κB1,轉(zhuǎn)化大腸埃希氏菌BL21(DE3)。經(jīng)酶切和測(cè)序鑒定正確后,異丙基硫代-β-D-半乳糖苷(IPTG)大量誘導(dǎo)表達(dá),8 mol·L~(-1)尿素裂解包涵體,裂解后上清經(jīng)GST-Sepharose 4B親和純化,SDS-PAGE和Western blotting鑒定。結(jié)果表明,原核表達(dá)載體p GEX-4T-1-NF-κB1構(gòu)建成功,SDS-PAGE證實(shí)NF-κB1融合蛋白存在于包涵體內(nèi),純化后得到GST-NF-κB1融合蛋白,為進(jìn)一步研究NF-κB1蛋白在奶牛泌乳中作用提供試驗(yàn)依據(jù)。
[Abstract]:NF- 魏 B1 is an important nuclear transcription factor. NF- 魏 B1 is involved in the regulation of gene expression. NF- 魏 B1 can bind to the genes related to the regulation of milk synthesis in milk of dairy cows and play a role in the regulation of lactation.The coding sequence of NF- 魏 B1 gene in dairy cattle was amplified by PCR method. The recombinant plasmid pGEX-4T-1-NF- 魏 B1 was constructed by inserting Eco R 鈪，

本文編號(hào)：1763190