Establishment of Stable Expression hTERT Testicular Primary Cell Line
The in vitro culture of primary sheep testicular cells is an important material to explore the pathogenic mechanism of virus infection of bovine and sheep.The actual scientific research work is often trapped by the shortcomings such as poor passage and unstable properties of these cells.Exogenous introduction of human telomerase reverse transcriptase gene(human telomerase reverse transcriptase gene,hTERT)can promote the expression of telomerase activity extend the lifespan of the cells in vitro,which is an effective method to establish cell immortalization.In this study,hTERT was amplified by RT-PCR and the eukaryotic expression plasmid and lentivirus plasmid of hTERT was constructed by homologous recombination technique.Then,the stable sheep testicular cell lines were established by using the lentivirus expression system.The results of indirect immunofluorescence and Western blotting showed that the hTERT gene had been integrated into the sheep testicular genome with stable expression.The sheep testicular cells of the 30th passage grew faster and their characters were relatively stable,which indicated that an immortalized sheep testicular cell line was established which could provide an important cell model for the related research of herbivore virus.
国家科技期刊平台
NSTL
万方数据
维普
