中国动物传染病学报2024，Vol.32Issue(2)：43-48,6.
稳转hTERT绵羊睾丸细胞系的建立
Establishment of Stable Expression hTERT Testicular Primary Cell Line
摘要
Abstract
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.关键词
绵羊/睾丸原代细胞/人端粒酶逆转录酶基因/hTERT/永生化细胞系Key words
Sheep/primary testicle cell/human telomerase catalytic subunit/hTERT/immortalized cell lines分类
农业科技引用本文复制引用
杨雪,任善会,王相伟,龚真莉,殷相平,孙跃峰,陈豪泰,万学瑞..稳转hTERT绵羊睾丸细胞系的建立[J].中国动物传染病学报,2024,32(2):43-48,6.基金项目
甘肃农业大学兽医学省级特色学科开放课题(GSAU-XKJS-2018-071) （GSAU-XKJS-2018-071）
中国农业科学院兰州兽医研究所基本科研业务费(1610312021008) （1610312021008）
