| 摘要: |
| 为了构建端粒酶逆转录酶表达载体，用带有绿色荧光蛋白报告基因的载体pEGFP-C1和带有hTERTcDNA的载体pCL-Neo-hTERT，通过基因重组构建新的载体pEGFP-hTERT，转化后筛选阳性克隆进行酶切鉴定；pEGFP-hTERT载体通过脂质体介导法转染纯化后的牛A型精原细胞，观察绿色荧光表达，G418筛选阳性克隆。结果表明，pEGFP-hTERT载体与预设计的一致；pEGFP-hTERT载体转染牛A型精原细胞后，经筛选获得了1个表达绿色荧光蛋白的阳性细胞克隆。由此可见，hTERT表达载体得到了成功构建，并在牛A型精原细胞中实现了表达。 |
| 关键词: A型精原细胞 基因重组 hTERT 牛 |
| DOI： |
| 分类号: |
| 基金项目:国家“863”计划项目（2004AA213072) |
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| Establishment of pEGFP-hTERT vector and expression in type A spermatogonia of bovine |
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| Abstract: |
| To construct a telomerase reverse transcriptase expression vector,a new vector pEGFP-hTERT was established by genetic recombination of vector pEGFP-C1 with green fluorescence protein reporter gene and vector pCL-Neo-hTERT containing hTERT cDNA.Positive clones containing recombinant plasmids were identified by restriction endonuclease digesting.Type A spermatogonia of bovine after purification were transfected with pEGFP-hTERT vector mediated by lipofectamine and the expression of green fluorescence protein was observed,and then the positive clone was screened with G418.The results showed that the pEGFP-hTERT vector was identical to the size devised in advance.After transfecting with pEGFP-hTERT vector,a positive clone of type A spermatogonia expressing green fluorescence protein of bovine was obtained by screening.Thus it could be concluded that pEGFP-hTERT vector was established and expressed in type A spermatogonia of bovine successfully. |
| Key words: type A spermatogonia gene recombination hTERT bovine |
