[Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell]

Yao Xue Xue Bao. 2005 Sep;40(9):810-3.
[Article in Chinese]

Abstract

Aim: To investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line.

Methods: The HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting.

Results: The recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The M(w) of the recombinant protein was about 120 kD.

Conclusion: Recombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3',5'-Cyclic-AMP Phosphodiesterases / genetics
  • 3',5'-Cyclic-AMP Phosphodiesterases / metabolism*
  • Animals
  • Baculoviridae / genetics*
  • Cell Line
  • Cyclic Nucleotide Phosphodiesterases, Type 3
  • Electrophoresis, Polyacrylamide Gel
  • Moths / cytology
  • Moths / enzymology*
  • Moths / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transfection

Substances

  • RNA, Messenger
  • Recombinant Proteins
  • 3',5'-Cyclic-AMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 3
  • PDE3A protein, human