文献类型：期刊文献

中文题名：菜豆晕疫病原菌甘油激酶基因敲除突变体的构建与表型分析

英文题名：Construction of glycerol kinase gene deletion mutants of Pseudomonas syringae pv. phaseolicola and its phenotype analysis

作者：刘晓柱 李银凤 于志海 刘晓辉 黄名正

第一作者：刘晓柱

机构：[1]贵州理工学院食品药品制造工程学院

第一机构：贵州理工学院食品药品制造工程学院

年份：2018

卷号：44

期号：2

起止页码：145-150

中文期刊名：湖南农业大学学报：自然科学版

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：贵州省教育厅创新群体重大研究项目(黔教合KY字[2017]046);贵州省理工学院混合教学模式课程建设项目(2017HHKC06)

语种：中文

中文关键词：菜豆晕疫病菌;甘油激酶基因;敲除突变体;表型分析

外文关键词：Pseudomonas syringae pv.phaseolicola;glycerol kinase gene;deletion mutants;phenotypical analysis

摘要：以菜豆晕疫病菌(Pseudomonas syringae pv.phaseolicola)NPS3121株系为出发材料,根据Gen Bank中登录的1448A甘油激酶(glycerol kinase,GK)氨基酸序列设计同源引物,通过PCR克隆NPS3121 GK基因,序列全长为1 506 bp,可编码1条含501个氨基酸的多肽。生物信息学分析表明,NPS3121 GK包含469个氨基酸,相对分子质量为55 800,富含α–螺旋结构,等电点为5.44。通过整合突变的方式构建了NPS3121 GK敲除突变体,敲除突变体与野生型相比,在M9培养基中生长速率降低18%,在KBM培养基中生长速率降低16%。敲除突变体中甘油浓度较野生型提高了6倍。
GK coding sequence of Pseudomonas syringae pv.phaseolicola strain NPS3121 was cloned by PCR with the primers designed according to reported 1448A glycerol kinase(GK)amino acid sequence in GenBank.Sequencing analysis showed that the length of the GK gene was 1 506 bp,which could encode a polypeptide containing 501 amino acids.Bio–informatics analysis indicated that GK of strain NPS3121 contained 469 amino acids,with molecular weight 55 800,enriched in alpha helix,and isoelectric point was 5.44.GK deletion mutants of strain NPS3121 were constructed by integrated mutation and the results showed that the growth rate of GK deletion mutants reduced 18%and 16%,respectively in M9 and KBM medium,compared with wild strain.In addition,the content of glycerol in GK deletion mutants increased 6 times,compared to the wild strain.