Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell.
- Author:
Yong-Min YAN
1
;
Wen-Rong XU
;
Hai-Bo ZHU
;
Ling ZHANG
;
Ren-Ren FEI
;
Xiao-Liang WANG
;
Hui QIAN
;
Shun-Zu HAO
;
Wen-Bing WANG
Author Information
- Publication Type:Journal Article
- MeSH: 3',5'-Cyclic-AMP Phosphodiesterases; genetics; metabolism; Animals; Baculoviridae; genetics; Cell Line; Cyclic Nucleotide Phosphodiesterases, Type 3; Electrophoresis, Polyacrylamide Gel; Moths; cytology; enzymology; metabolism; RNA, Messenger; genetics; metabolism; Recombinant Proteins; genetics; metabolism; Transfection
- From: Acta Pharmaceutica Sinica 2005;40(9):810-813
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line.
METHODSThe 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.
RESULTSThe 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.
CONCLUSIONRecombinant 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.
