Cloning and expression of apolipoprotein E3 and its variant apoE2 and apoE4
Zong Yiqiang , Liu Zhiguo , Bi Hao , Yao Yanyi , Guo Jianli , Qu Shen
Current Medical Science ›› 2006, Vol. 26 ›› Issue (1) : 1 -3.
Cloning and expression of apolipoprotein E3 and its variant apoE2 and apoE4
In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.
apoE / gone cloning / site-directed mutation / gene expression
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