奶牛载脂蛋白B100基因克隆及原核表达

Abstract

Abstract: Specific primers were designed according to ApoB100 gene sequence reported in GenBank in this study. Cloning was carried out by the pGM-T vector, recombined plasmid DNA was cut by BamHⅠand EcoRⅠenzymes and then sequencing. The recombinant expression vector pET-28a-ApoB100 was constructed with target gene and pET-28a vector, and transformed into Rosetta(DE3),then induced by IPTG. The expression product was identified by SDS-PAGE and Western blotting. The results showed that the homology of the cloned ApoB100 gene was 100% to that reported in GenBank. SDS-PAGE analysis showed that the molecular weight of recombinant protein pET-28a-ApoB100 was 35 ku. Western blotting positive results showed that the experiment successfully obtained the target protein.

Key words: ApoB100; gene cloning; prokaryotic expression

CLC Number:

Q786

ZHANG Jia-li, WANG Li, YANG Wen-tao, DING Hong-yan, ZHANG Liang, CHEN Hui, LIU Zhao-xi, ZHAO Chen-xu, WANG Zhe, LI Xiao-bing. Cloning and Prokaryotic Expression of Apolipoprotein B100 Gene of Dairy Cows[J]. , 2012, 39(11): 43-46.

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Cloning and Prokaryotic Expression of Apolipoprotein B100 Gene of Dairy Cows

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