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Cloning and expression of apolipoprotein E3 and its variant apoE2 and apoE4

  • Zong Yiqiang
  • Liu Zhiguo
  • Bi Hao
  • Yao Yanyi
  • Guo Jianli
  • Qu Shen
Article

Summary

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.

Key words

apoE gone cloning site-directed mutation gene expression 

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Copyright information

© Springer 2006

Authors and Affiliations

  • Zong Yiqiang
    • 1
  • Liu Zhiguo
    • 2
  • Bi Hao
    • 1
  • Yao Yanyi
    • 1
  • Guo Jianli
    • 1
  • Qu Shen
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Department of Biotechnology and Chemical EngineeringWuhan Polytechnic UniversityWuhanChina

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