沧州沿海湿地盐地碱蓬P5CS基因及启动子克隆研究  

Cloning of P5CS gene and promoter from Suaeda salsa in Cangzhou coastal wetlands

作　　者：王君 田景汉 彭惠 WANG Jun;TIAN Jing-han;PENG Hui

机构地区：[1]沧州师范学院生命科学学院,河北沧州061001

出　　处：《饲料研究》2020年第7期85-89,共5页Feed Research

基　　金：河北省科技支撑计划项目(项目编号:16273306);河北省高等学校科学技术研究项目(项目编号:2020103021424);沧州市科学技术研究与发展重点研发计划资助项目(项目编号:192203003);沧州师范学院滨海环境生态创新团队资助(项目编号:cxtdl1904)。

摘　　要：为深入研究盐地饲用植物碱蓬(Suaeda salsa)Δ1-吡咯啉-5-羧酸合成酶(P5CS)基因的功能,在已知盐地碱蓬P5CS基因c DNA序列的基础上,对耐盐P5C5基因组进行克隆及分析。选用TAIL-PCR的试验方法对其上游未知序列进行探索,并对其编码的P5CS蛋白及相关植物P5CS基因编码的蛋白进行生物信息学分析。获得该基因序列片段长为1506 bp。与已知的c DNA序列进行同源性比对,对比结果表明试验所获得的序列片段为盐地碱蓬P5CS基因组序列。通过与NCBI网站的Proteins数据库中相关植物P5CS蛋白序列进行比对,进化分析表明盐地碱蓬P5CS与北美海蓬子的亲缘关系最近,氨基酸序列相似度达91%,同时在盐地碱蓬P5CS基因的启动子克隆中筛选了40种RAPD引物,为进一步研究耐盐基因启动子克隆及功能表达奠定基础,对盐地饲用植物碱蓬耐盐的分子机理研究提供参考依据。In order to study the function ofΔ1-pyrrolidine-5-carboxylic acid synthase(P5 CS)gene of Suaeda salsa,the salt-tolerant P5 C5 genome was cloned and analyzed on the basis of the known P5 CS gene sequence.TAIL-PCR experimental method was selected to explore its upstream and unknown sequence,its genome was cloned and analyzed.Finally,proteins encoded by Suaeda salsa and related plants P5 CS genes were analyzed in bioinformatics.The obtained sequence fragment is 1506 bp in length.Compared with the known c DNA sequence homologously,the comparison result showed that the obtained sequence fragment was the Suaeda salsa P5 CS genomic sequence.By comparing with the P5 CS protein sequence of related plants in the proteins database of NCBI website,the phylogenetic analysis showed that the genetic relationship between Suaeda salsa P5 CS and Salicornia hainanensis was the closest,and the similarity of amino acid sequence was 91%.At the same time,forty RAPD primers were screened from the promoter clones of Suaeda salsa P5 CS gene,the study laid the foundation for the promoter cloning and functional gene expression in the subsequent experiments,and also provided basis for the study on the molecular mechanism of salt-tolerance in Suaeda salsa.

关 键 词：P5CS TAIL-PCR 启动子基因组 

分 类 号：S816[农业科学—饲料科学]