The Chinese-German Journal of Clinical Oncology

, Volume 5, Issue 5, pp 358–361

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

Authors

  • Wenhua Xiong
    • Department of Orthopedics, Tongji HospitalHuazhong University of Science and Technology
  • Anmin Chen
    • Department of Orthopedics, Tongji HospitalHuazhong University of Science and Technology
  • Fengjing Guo
    • Department of Orthopedics, Tongji HospitalHuazhong University of Science and Technology
  • Tao Huang
    • Department of Orthopedics, Tongji HospitalHuazhong University of Science and Technology
Article

DOI: 10.1007/s10330-005-0450-1

Cite this article as:
Xiong, W., Chen, A., Guo, F. et al. Chinese German J Clin Oncol (2006) 5: 358. doi:10.1007/s10330-005-0450-1
  • 85 Views

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

Pin1isomeraseantisense geneeukaryotic expressing vector
Download to read the full article text

Copyright information

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