Cloning and Expression of Apolipoprotein E3 and Its Variant apoE2 and apoE4

In order to obtain three isoforms of apolipoprotein E （apoE）, the eDNA 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 se...

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Bibliographic Details
Published in:Journal of Huazhong University of Science and Technology. Medical sciences 2006, Vol.26 (1), p.1-3
Main Author: 宗义强 刘志国 毕昊 姚研怡 过健俐 屈伸
Format: Article
Language:English
Subjects:
apoE2
apoE4
Apolipoprotein E2
Apolipoprotein E3
Apolipoprotein E4
Apolipoproteins E - biosynthesis
Apolipoproteins E - genetics
Apolipoproteins E - isolation & purification
Base Sequence
Cloning, Molecular
Humans
Liver - chemistry
Molecular Sequence Data
Point Mutation
变异体
基因表达
无性繁殖
载脂蛋白E3
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Summary:In order to obtain three isoforms of apolipoprotein E （apoE）, the eDNA 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 eDNA into vector pTT-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.
ISSN:1672-0733
1993-1352
DOI:10.1007/bf02828023