西安交通大学学报（医学版）2013，Vol.34Issue(2)：263-266,4.
pEGFP-N1-TNFα表达载体的构建与鉴定
Construction and identification of pEGFP-N1-TNFα expression vector
摘要
Abstract
Objective To construct and indentify a recombinant pEGFP-Nl-TNFa expression plasmid so as to lay a foundation for transfecting CIK cells and localize the expression. Methods pMD19 Simple T-TNFa was taken as template, the fragment Supported by the Technological Plan of Social Development of Yunnan Province (Fundamental Research Program, No. 2009ZC119M), the Health and Science Research Institutes Foundation of Yunnan Province (No. 2011WS0068), and the Technological Plan of Social Development of Yunnan Province (Key Fundamental Research Program, No. 2009CC026) of CDS region of TNFa was amplified by PCR, and then the products of PCR amplification underwent agarose gel electrophoresis. The fragment TNFa was excised, and then fragment TNFa and aimed vector were excised by Bgl Ⅱ and Hind Ⅲ double digestion. The excised products were connected; the product was then transformed into DH5a. The positive clones were selected, from which plasmid DNA was abstracted and identified by sequencing. Results The size of PCR products and double digested section were 722 bp, which was in accordance with the expected results; sequence analysis of inserted fragment revealed the same sequence as synthesize oligonucleotides. Conclusion The pEGFP-Nl-TNFa expression vector had been successfully constructed, which laid a foundation for future study on nanomaterial-mediated transfection into CIK cells.关键词
肿瘤坏死因子/真核表达载体/质粒pEGFP-N1/基因疗法/CIK细胞Key words
tumor necrosis factor (TNF)/ eukaryocyte expression vector/ pEGFP-N1 plasmid/ gene therapy/ CIK cell分类
医药卫生引用本文复制引用
任艳鑫,赵留芳,杨洁,孙瑞梅,王虎,李晓江,隋军..pEGFP-N1-TNFα表达载体的构建与鉴定[J].西安交通大学学报（医学版）,2013,34(2):263-266,4.基金项目
云南省社会发展科技计划(应用基础研究项目,No.2009ZC119M) （应用基础研究项目,No.2009ZC119M）
云南省卫生科技内设研究机构资助项目(No.2011WS0068) （No.2011WS0068）
云南省社会发展科技计划(应用基础研究重点项目,No.2009CC026) （应用基础研究重点项目,No.2009CC026）
