殊异韦荣菌乳酸脱氢酶基因的克隆及鉴定  

Cloning and identification of lactate dehydrogenase gene of Veillonella dispar

作　　者：庾桦[1,2] 何钟勤[3] 高心[1] 钟丞[1] 李倪娜[1] 薛莹[1] 王志刚[4] 

机构地区：[1]吉林大学口腔医院牙体牙髓病科,吉林长春130021 [2]承德医学院附属医院口腔内科,河北承德067000 [3]吉林大学中日联谊医院口腔科,吉林长春130033 [4]中国石油吉化集团公司总医院口腔科,吉林吉林132022

出　　处：《吉林大学学报（医学版）》2011年第5期825-828,共4页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(201015204)

摘　　要：目的:对殊异韦荣菌(V.dispar)乳酸脱氢酶(LDH)基因及其前后基因进行克隆和鉴定,为龋病的预防和治疗奠定理论基础。方法:提取殊异韦荣菌基因组DNA,利用PCR方法扩增LDH及其前后同源区序列片段,并克隆入pMD18-T载体中,转化大肠杆菌JM109,酶切及PCR鉴定后,进行测序。结果:PCR扩增产物特异,全长2 300 bp。测序结果经BLAST软件分析,共包含3个基因,分别是DNA旋转酶(gyr A)、LDH和一个由未知蛋白编码的基因。并与GenBank中所报道的变形链球菌LDH基因序列进行对比分析,其同源性为99%,该基因序列已登陆GenBank,序列号为EU518464。结论:成功克隆殊异韦荣菌LDH基因,并通过基因序列和氨基酸分析证明其具有完整的阅读框架。Objective To clone and detect the lactate dehydrogenase（LDH） gene of Veillonella dispar（V.dispar） and its adjacent gene and provide a theoretical basis for prevention and treatment of dental caries.Methods LDH gene and its adjacent gene were detected by PCR method and were inserted into a cloning vector PMD18-T to transform E.coil JM109.The sequencing was performed after PCR identification and restriction enzyme digestion.Results The PCR product was specific,and the size of the product was 2 300 bp.The sequencing data were analyzed by BLAST software and the result indicated that they contained three pieces of genes which were DNA gyrase A,LDH,and an unknown product.The homology of the sequencing result was 99 % compared with the LDH gene of Streptococcus（S.mutans） which was reported in GenBank.And the sequence had been submitted to the NCBI GenBank,and the accession number was EU518464.Conclusion The LDH gene of Veillonella dispar was successfully cloned,and it is proved to have a full reading framework,through the analysis of the gene sequences and amino acids.

关 键 词：殊异韦荣菌 乳酸脱氢酶 基因克隆 序列测定 

分 类 号：R781.1[医药卫生—口腔医学] Q78[医药卫生—临床医学]