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


CONCLUSIONS
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.
Yan
Yong-Min
YM
School of Medical Technology, Jiangsu University, Zhenjiang 212001, China.
Xu
Wen-Rong
WR
Zhu
Hai-Bo
HB
Zhang
Ling
L
Fei
Ren-Ren
RR
Wang
Xiao-Liang
XL
Qian
Hui
H
Hao
Shun-Zu
SZ
Wang
Wen-Bing
WB
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; D004591:Electrophoresis, Polyacrylamide Gel; D009036:Moths / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D012333:RNA, Messenger; D011994:Recombinant Proteins; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; C517287:PDE3A protein, human