Cloning and construction of sense and antisense eukaryotic expression vector of human Pin1

  • Wenhua Xiong
  • Anmin Chen
  • Fengjing Guo
  • Tao Huang
Article
  • 95 Downloads

Abstract

Objective: To clone and construct eukaryotic expressing vectors of sense and antisense human Pin1 (hPin1) genes. Methods: Total RNA was extracted from MG-63 cells, then the hPin1 cDNA was amplified by RT-PCR. The same time the sense and antisense hPin1 genes were formed by binding BamH I and Hind III in cis and trans-directions. At the end they were cloned into the eukaryotic expressing vector pIRES2-EGFP in cis and trans directions using DNA recombinant technology. The recombinant vectors were further identified by digestion of BamH I and Hind III. Results: The results of sequencing showed that the orientation of the ligations and the reading frame were correct. After digested by BamH I and Hind III, two fragments exhibiting 5.3 kb and 0.99 kb were formed in sense and antisense eukaryotic expressing vectors. Electrophoretic results were completely coincident with theoretical calculation. Conclusion: Human Pin1 sense and antisense genes were successfully cloned and eukaryotic expressing vectors were successfully constructed.

Key words

Pin1 isomerase antisense gene eukaryotic expressing vector 

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Copyright information

© Editorial Office of the Chinese-German Journal of Clinical Oncology 2006

Authors and Affiliations

  • Wenhua Xiong
    • 1
  • Anmin Chen
    • 1
  • Fengjing Guo
    • 1
  • Tao Huang
    • 1
  1. 1.Department of Orthopedics, Tongji HospitalHuazhong University of Science and TechnologyWuhanChina

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