广西陆川猪肺炎支原体P46基因克隆及序列分析  

Cloning and Sequence Analysis of P46 Gene of Mycoplasma hyopneumoniaein Guangxi Luchuan Pig

作　　者：李莹莹[1,2] 赵武[1] 李斌[1] 秦毅斌[1] 卢冰霞[1] 梁家幸[1] 何颖[1] 周英宁[1] 陆芹章[2] 

机构地区：[1]广西兽医研究所广西畜禽疫苗新技术重点实验室,广西南宁530001 [2]广西大学动物科学技术学院,广西南宁530005

出　　处：《中国畜牧兽医》2014年第2期1-5,共5页China Animal Husbandry & Veterinary Medicine

基　　金：广西水产畜牧兽医局科研计划(桂渔牧科1304522、1204929、1204931);广西畜禽疫苗新技术重点实验室专项(12-071-28-A-5);广西科技攻关项目(桂科攻1123007-3)

摘　　要：为了解广西陆川猪肺炎支原体（Mhp）地方流行毒株的主要免疫原性表面蛋白基因P46的序列特征、基本结构及遗传变异等特点，试验以2011至2013年广西纯种陆川猪疑似猪支原体肺炎（MPS）阳性病料为研究对象，抽提DNA进行PCR扩增，将克隆出与目的基因大小相符的片段回收，再与载体连接，转化到大肠杆菌DH5“感受态细胞中，筛选阳性克隆及测序，扩增获得4株Mhp毒株（GXLC1、GXLC2、GXLC3和GXLC4），随后用DNAStar软件对P46基因序列进行比对分析。结果表明，已克隆出的P46基因序列长度为1104bp，编码368个氨基酸，其中该序列中含有3个编码色氨酸的TGA密码子，而不是终止密码子；1个CGG为编码精氨酸的稀有密码子，而不是其他支原体中的无义密码子。所获得的4株Mhp毒株的P46基因序列与所参考的MhpJ株、232株、7448株及168株的核苷酸同源性为98．4％～99．4％，氨基酸同源性为98．6％～99．5％，毒株之间P46基因的同源性很高，保守性较强。In order to understand the sequence characteristics, basic structure and genetic variation of P46 gene which was the main immunogenic surface membrane protein gene of local prevalent Mycoplasma hyopneumoniae （Mhp） strains in Guangxi Luchuan pig. The Mhp P46 gene in positive diseased swine samples which were collected from the purebred Luchuan pig farms in Guangxi from 2011 to 2013 was amplified using PCR, and then cloned into pMD18-T vector and transformed into E. coli DHSa. We chose the positive clones and sequenced. We amplified P46 genes of four positive strains （GXLC1, GXLC2,GXLC3 and GXLC4）. Use the DNAStar software to analyse the cloned sequence. The results showed that P46 gene sequence was 1104 bp coding 368 amino acids. The sequence included three Trps coded by TGA codons which weren＇t termination codons and one Arg coded by CGG codons which weren＇t nonsense code. The homologies of nucleotide sequences of 4 strains were 98.4% to 99.4% ,and the homologies of deduced amino acid sequences were 98.6% to 99.5% with these sequences of standard J strain,232 strain,7448 strain and 168 strain. The P46 genes of these strains had highly conservation because of the high-level homologies.

关 键 词：广西陆川猪 猪肺炎支原体 P46基因 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]