| 刘转转1 郝林2 范涛2 贺厚光2 尤红娟1 汤仁仙1 韩从辉2.人TRAIL 基因原核表达载体的构建及鉴定[J].现代生物医学进展英文版,2011,11(7):1220-1223. |
| 人TRAIL 基因原核表达载体的构建及鉴定 |
| Construction and Identification of Prokaryotic Expression Vector of HumanTNF-related Apoptosis Inducing Ligand |
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| DOI： |
| 中文关键词: TRAIL RT-PCR 克隆 |
| 英文关键词: TRAIL RT-PCR Cloning |
| 基金项目:国家自然科学基金项目（30973443），徐州市科技计划项目（XM08C055） |
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| 中文摘要: |
| 目的：克隆肿瘤坏死因子相关凋亡诱导配体（TRAIL）基因片段（114～281 氨基酸残基）并构建原核表达载体。方法：取健康
人外周血提取总RNA，设计合成引物并引入EcoR I 和Xho I 酶切位点，RT-PCR 扩增TRAIL 基因的胞外区片段，克隆入原核表
达载体pGEX-6P-1 中，经双酶切、PCR 及测序鉴定阳性克隆。结果：从外周血cDNA 中扩增出501 bp 的目的片段，测序结果证实
成功构建重组质粒pGEX-6P-1 /TRAIL。结论：成功构建TRAIL 基因的原核表达载体pGEX-6P-1 /TRAIL，为肿瘤细胞的凋亡研究
提供理论依据。 |
| 英文摘要: |
| Objective: To clone and construct the prokaryotic expression vector of the TNF-related apoptosis inducing ligand gene
（amino acids 114～281）. Methods: RNA was extracted from the peripheral blood of health adult. A pair of primers with restriction site
EcoR I/Xho I was designed. The extracellular domain region of TRAIL gene was amplified by RT-PCR and then cloned into the prokaryotic
expression vector pGEX-6P-1. The recombinants were confirmed by EcoR I/Xho I digestion, PCR, and DNA sequencing. Result:
The extracellular domain region with a molecular size of 501 bp of TRAIL gene was amplified successfully. DNA sequencing showed
that the recombinant plasmid was successfully constructed. Conclusion: A prokaryotic expression vector pGEX-6P-1/TRAIL was constructed
successfully, which provides theory for tumor apoptosis research. |
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