›› 2010, Vol. 37 ›› Issue (9): 70-74.

• 生物技术 • Previous Articles     Next Articles

Prokaryotic Expression of Porcine Phosphodiesterase 4B2(PDE4B2and its Activity

CHEN Li1, JIANG Dai-xun2, REN Chao1, CHEN Yi-shan1, LI Jia2, CHEN Wu1   

  1. (1.College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China;2.Beijing Key Laboratory of Veterinary Medicine (Traditional Chinese Medicine and Pharmacology), Beijing 102206, China）
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-09-20 Published:2010-09-20
  • Contact: CHEN Wu

Abstract: Porcine phosphodiesterase 4B2 gene was amplified by RT-PCR method and cloned into the prokaryotic expression vector. The positive recombinant plasmid, which was screened by PCR and sequencing, was transformed into E.coli strain BL21（DE3） and recombinant protein expression was induced with IPTG. The results showed that the target fragment was 1718 bp in length and has 99.7% homology with that of porcine counterparts deposited in GenBank. The protein was produced in inclusion bodies and soluble forms. The recombination PDE4B2 protein was 66 ku in molecular mass by Western blotting analysis. Activity of the cAMP-PDE was 51.46%, detected by high performance liquid chromatography (HPLC). The polyclonal antibody raised against the recombinant PDE4B2 protein had a good specificity, which lays a foundation for nature PDE4B2 protein and inhibitor of PDE4.

Key words: porcine phosphodiesterase 4B2; prokaryotic expression; Western blotting; polyclonal antibody

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