本文選題：奶牛 + NF-κB　； 參考：《東北農(nóng)業(yè)大學學報》2016年09期

【摘要】：NF-κB1是一類重要核轉錄因子,參與多種基因表達調(diào)控。NF-κB1能與奶牛乳腺中乳合成調(diào)控相關基因結合,發(fā)揮泌乳調(diào)節(jié)作用。研究應用PCR方法擴增奶牛NF-κB1基因編碼序列,通過Eco RⅠ和XhoⅠ酶切位點將其定向插入p GEX-4T-1載體中,構建原核表達重組質(zhì)粒p GEX-4T-1-NF-κB1,轉化大腸埃希氏菌BL21(DE3)。經(jīng)酶切和測序鑒定正確后,異丙基硫代-β-D-半乳糖苷(IPTG)大量誘導表達,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。結果表明,原核表達載體p GEX-4T-1-NF-κB1構建成功,SDS-PAGE證實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,為進一步研究NF-κB1蛋白在奶牛泌乳中作用提供試驗依據(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：1763190