无乳链球菌GlnR因子DNA结合位点预测及其突变基因的构建  

Prediction of DNA-binding Sites in Streptococcus agalactiae GlnR Factor and Construction of glnR Mutant Gene

作　　者：夏杰[1] 易弋[1] 王佳[1] 杨军 黄稀 黄翠姬[1] 伍时华[1] 

机构地区：[1]广西工学院,广西柳州545006 [2]柳州市渔业技术推广站,广西柳州545006 [3]柳州市鱼家乐饲料有限公司,广西柳州545002

出　　处：《湖北农业科学》2013年第23期5902-5905,共4页Hubei Agricultural Sciences

基　　金：广西教育厅科研项目(201010LX217);广西科技大学(筹)科学基金资助项目(校科自1261125)

摘　　要：利用生物信息学的方法对无乳链球菌（Streptococcus agalactiae）GlnR因子与DNA结合的位点进行预测,筛选出GlnR因子中与DNA结合的关键位点的氨基酸残基R47和R48.然后利用重叠PCR的方法将glnR基因上的第139～144位上的碱基CGCCGC突变为GCGGCG,使其编码的2个精氨酸残基突变为丙氨酸残基,将突变基因片段TA克隆至PMD 18-T载体上进行测序验证.测序结果表明,利用重叠PCR成功突变了这6个碱基,为后续研究GlnR因子的突变位点对DNA结合的影响提供了参考.Using bioinformatics methods,the key DNA binding sites of amino acid residues R47 and R48 were screened out in GlnR factor by predicting DNA binding sites of the Streptococcus agalactiae GlnR factor.The overlap PCR method was used to mutate the glnR gene from CGCCGC to GCGGCG at 139th to 144th sites,namely altering two coded arginine residues to alanine residues.The mutant gene fragment TA was cloned into PMD18-T vector and the sequence was validated.It showed that 6 bases was successfully mutated using the overlap PCR method and laid the basis for further functional analysis of the mutational site in GlnR factor for DNA binding.

关 键 词：无乳链球菌(Streptococcus agalactiae) glnR DNA结合位点 重叠PCR 生物信息学 

分 类 号：R378.12[医药卫生—病原生物学] Q-33[医药卫生—基础医学]