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

  • Wenhua Xiong
  • Anmin Chen
  • Fengjing Guo
  • Tao Huang


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 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Blume-Jensen P, Hunter T. Oncogenic kinase signalling. Nature, 2001, 411: 355–365.PubMedCrossRefGoogle Scholar
  2. 2.
    Hanahan D, Weinberg RA. The hallmarks of cancer. Cell, 2000, 100: 57–70.PubMedCrossRefGoogle Scholar
  3. 3.
    Lu KP, Liou YC, Zhou XZ. Pinning down the praline-directed phosphorylation signalling. Trends Cell Biol, 2002, 12: 164–172.PubMedCrossRefGoogle Scholar
  4. 4.
    Ryo A, Liou YC, Lu KP, et al. Prolyl isomerase Pin1: a catalyst for oncogenesis and a potential therapeutic target in cancer. J Cell Sci, 2003, 116: 773–783.PubMedCrossRefGoogle Scholar
  5. 5.
    Bao L, Kimzev A, Sauter G, et al. Prevalent overexpression of proly1 isomerase Pin1 in human cancer. Am J Pathol, 2004, 164: 1727–1737.PubMedGoogle Scholar
  6. 6.
    Nakamura M, Ryo A, Wulf G, et al. Pin1 is an E2F target gene essential for Neu/Ras-induced transformation of mammary epithelial cells. Mol Cell Biol, 2002, 22: 5281–5295.PubMedCrossRefGoogle Scholar
  7. 7.
    Weinberg RA. The molecular basis of oncogenes and tumor suppressor genes. Ann NY Acad Sci, 1995, 758: 331–338.PubMedGoogle Scholar
  8. 8.
    Nevins JR. The Rb/E2F pathway and cancer. Hum Mol Genet, 2001, 10: 699–703.PubMedCrossRefGoogle Scholar
  9. 9.
    Ryo A, Nakamura N, Wulf G. Pin1 regulates turnover and subcellular localization of beta-catenin by inhibiting its interaction with APC. Nat Cell Biol, 2001, 3: 793–801.PubMedCrossRefGoogle Scholar
  10. 10.
    Liou YC, Ryo A, Huang HK, et al. Loss of Pin1 function in the mouse causes phenotypes resembling cyclin D1-null phenotypes. Proc Natl Acad Sci USA, 2002, 99: 1335–1340.PubMedCrossRefGoogle Scholar
  11. 11.
    Ayala G, Wang D, Wulf G, et al. The prolyl isomerase Pin1 is a novel prognostic marker in human prostate cancer. Cancer Res, 2003, 63: 6244–6251.PubMedGoogle Scholar
  12. 12.
    Lu PJ, Zhou XZ, Shen M, et al. Function of WW domains as phosphoserine-or phosphothreonine-binding modules. Science, 1999, 283: 1325–1328.PubMedCrossRefGoogle Scholar
  13. 13.
    Wulf GM, Liou YC, Ryo A, et al. Role of Pin1 in the regulation of p53 stability and p21 transactiviation, and cell cycle checkpoints in response to DNA damage. J Biol Chem, 2002, 277: 47976–47979.PubMedCrossRefGoogle Scholar
  14. 14.
    Rippmann JF, Hobbie S, Daiber C, et al. Phosphorylation-dependent proline isomerization catalyzed by Pin1 is essential for tumor cell survival and entry into mitosis. Cell Growth Differ, 2000, 11: 409–416.PubMedGoogle Scholar
  15. 15.
    Fujimori F, Takahashi K, Uchida C, et al. Mice lacking Pin1 develop normally, but are defective in entering cell cycle from G(0) arrest. Biochem Biophys Res Commun, 1999, 265: 658–663.PubMedCrossRefGoogle Scholar

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

Personalised recommendations