Cloning and Expression of Apolipoprotein E3 and Its Variant apoE2 and apoE4
- Author:
Yiqiang ZONG
1
;
Zhiguo LIU
;
Hao BI
;
Yanyi YAO
;
Jianli GUO
;
Shen QU
Author Information
1. 华中科技大学同济医学院
- Keywords:
apoE;
gene cloning;
site-directed mutation;
gene expression
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2006;26(1):1-3,16
- CountryChina
- Language:Chinese
-
Abstract:
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.
