重庆医学2017，Vol.46Issue(22)：3033-3035,3.DOI:10.3969/j.issn.1671-8348.2017.22.003
小鼠pMSV-Slfn5-GFP表达质粒构建及基因结构分析
Construction of pMSV-Slfn5-GFP plasmid and analysis of gene structure in mice
摘要
Abstract
Objective To perform mouse pMSV-Slfn5-GFP gene recombinant expression plasmid construction and gene structure analysis.Methods Total RNA was extracted from mouse liver and turned into cDNA by reverse transcription.Mouse Slfn5 coding sequence (CDS) fragment was amplified by PCR and connected with the pGEM-T Easy vector.The connected product was transferred the E.coli DH5a.The positive clones were selected for extracting plasmid,which was identified by double enzyme of restriction endonuclease Hpa Ⅰ and Xho Ⅰ.Then correct plasmid identified by enzyme digestion was sequenced by Macrogen USA.Then correct plasmid by sequencing was connected with pMSV-GFP by HindⅢ and Xbo Ⅰ,which was named as pMSV-Slfn5-GF.UCSC (http://genome.ucsc.Edu/) was used to analyze mouse Slfn5 and its family genomic structure.Slfn5 protein structural domain was determined by NCBI.Results Slfn5 full-length gene sequence was cloned into the expression vector pMSV-GFP,the fragment size was about 2.65 kb by enzyme digestion identification.The conservatism of AAA_4 protein domain in Slfn5 protein family was determined by phylogeny.fr.Conclusion Mouse full-length gene pMSV-Slfn5-GFP expression vector is successfully constructed.关键词
pMSV-Slfn5-GFP/构建/基因结构/分析Key words
pMSV-Slfn5-GFP/construction/gene structure/analysis分类
生物科学引用本文复制引用
况春燕,杨藩..小鼠pMSV-Slfn5-GFP表达质粒构建及基因结构分析[J].重庆医学,2017,46(22):3033-3035,3.基金项目
国家自然科学基金资助项目(81560056). （81560056）
