[The construction of pSecTag2/B vector modified by osteoprotegerin].
OBJECTIVE
In order to treat periodontitis by using tissue engineering and gene engineering technology, we constructed mammalian secreted expression pSecTag2/B-OPG vector, the cDNA sequence encoding osteoprotegerin(OPG) obtaining from 293 cell.


METHODS
The primers were designed based on the human OPG cDNA sequence. Total mRNA was isolated from 293 cells and RT-PCR was performed .The fragment was recombined into pSecTag2/B vector and sequenced by automatic sequence analyzer.


RESULTS
The sequences of OPG cDNA from 293 cell by RT-PCR were completely identical to the sequences provided by GenBank [gi:33878056]. After polymerase chain reaction and the recombinant plasmid digesting with Hind III,EcoR I and BamH I,1% agarose electrophoresis showed several fragments, which were consistent with predicted size.


CONCLUSIONS
From cultured 293 cells the cDNA has been cloned and pSecTag2/B-OPG vector has been constructed successfully. Supported by Research Fund (No.03043304) from Anhui Provincial Natural Science Foundation.
Department of Endodontics, College of Stomatology, Anhui Medical University, Hefei 230032, Anhui Province, China.
Yin
Y
Cao
Ling-xuan
LX
Mei
Di
D
Wu
Chun-hui
CH
Zhao
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D018076:DNA, Complementary; D005822:Genetic Vectors; D006801:Humans; D053244:Osteoprotegerin; D012333:RNA, Messenger
D018076:DNA, Complementary; D053244:Osteoprotegerin; D012333:RNA, Messenger