中国组织工程研究与临床康复2011，Vol.15Issue(24)：4489-4492,4.DOI:10.3969/j.issn.1673-8225.2011.24.029
分化型胚胎软骨发育基因1特异性小干扰RNA表达质粒的构建和鉴定
Construction and identification of small interfering RNA expression plasmid target to differentiated embryo-chondrocyte expressed gene 1
摘要
Abstract
BACKGROUND: Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is a basic helix-loop-helix transcription factor,which is closely associated with some malignant cancers.OBJECTIVE: To construct small interfering RNA (siRNA) expression plasmid target to DEC1.METHODS: The mRNA sequence of DEC1 gene was searched from NCBI. Utilize of Katahdin siRNA technology, DEC1-siRNA oligonucletides were inserted into pGreenPuro? shRNA Cloning and Expression Lentivector, after annealing, then transformed into JM-109. The recombinant plasmid was identified by Agarose gel electrophoresis analysis, ultraviolet spectrophotometer analysis, PCR and DNA sequencing.RESULTS AND CONCLUSION: The recombinant plasmid pGreenPuro? shRNA Cloning and Expression Lentivector-DEC1 was obtained by connecting 25 bp segment containing DEC1 sequence to pGreenPuro? shRNA Cloning and Expression Lentivector. Agarose gel electrophorsis analysis and ultraviolet spectrophotometer analysis confirmed that plasmid DNA had higher purity. DNA sequencing showed Targeting siRNA oligonucleotides were correctly inserted into the eukaryotic expression vector pGreenPuro? shRNA Cloning and Expression Lentivector without base mutation. The interference vector pGreenPuro? shRNA cloning and expression lentivector-DEC1 was successfully constructed.关键词
分化型胚胎软骨基因1/小干扰/RNA/构建/质粒/基因表达分类
医药卫生引用本文复制引用
魏炎,郏雁飞,郑燕,马晓丽,汪运山..分化型胚胎软骨发育基因1特异性小干扰RNA表达质粒的构建和鉴定[J].中国组织工程研究与临床康复,2011,15(24):4489-4492,4.基金项目
济南市科技局科技计划项目(200905045-1),国家自然科学基金(81000869)资助项目. （200905045-1）
