广西陆川猪猪肺炎支原体P97基因R1区克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2015.09.008

《中国畜牧兽医》---唯一指定的官方网站 ›› 2015, Vol. 42 ›› Issue (9): 2270-2277.doi: 10.16431/j.cnki.1671-7236.2015.09.008

广西陆川猪猪肺炎支原体P97基因R1区克隆及序列分析

黎珂钰^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, 韦英益²

1. 广西兽医研究所, 广西畜禽疫苗新技术重点实验室, 南宁 530001;
2. 广西大学动物科学技术学院, 南宁 530004

收稿日期:2015-01-26 出版日期:2015-09-20 发布日期:2015-09-25
通讯作者: 何颖, 韦英益 E-mail:heying921@163.com;weiyingyi99@163.com
作者简介:黎珂钰(1991-),女,广西百色人,硕士生,研究方向:动物病理学,E-mail:guucat@163.com
基金资助:
广西水产畜牧兽医局科研计划(桂渔牧科1304522、1204929);广西畜禽疫苗新技术重点实验室开发基金项目(14-045-31-B-3);南宁市西乡塘区科学研究与技术开发计划项目(2014304)

Cloning and Sequence Analysis of P97 Gene R1 Region of Mycoplasma hyopneumoniae in Guangxi Luchuan Pig

LI Ke-yu^1,2, ZHAO Wu¹, LI Bin¹, QIN Yi-bin¹, LU Bing-xia¹, LIANG Jia-xing¹, HE Ying¹, ZHOU Ying-ning¹, WEI Ying-yi²

1. Guangxi Key Laboratory of Animal Vaccines and New Technology, Guangxi Veterinary Research Institute, Nanning 530001, China;
2. College of Animal Science and Technology, Guangxi University, Nanning 530004, China

Received:2015-01-26 Online:2015-09-20 Published:2015-09-25

摘要/Abstract

摘要： 本试验通过深入了解广西陆川猪猪肺炎支原体(Mhp)地方流行毒株的黏附因子P97基因R1区的序列特征、基本结构及遗传变异等特点,为广西地方品种陆川猪猪支原体肺炎(MPS)有效的综合防控措施提供理论依据,并为进一步研究广西陆川猪Mhp地方流行毒株特性打下坚实的基础。参考Mhp基因组序列设计扩增P97基因R1区的1对引物,以 2011年—2014年广西陆川猪MPS阳性病料为研究对象,提取基因组DNA,PCR扩增P97基因R1区,对PCR产物进行测序,将广西陆川猪Mhp不同地方流行毒株的P97基因R1区序列的碱基组成和推导的氨基酸序列进行比对。DNAStar软件分析结果表明,所获得的15株广西陆川猪Mhp毒株(GXLC-1、GXLC-2、GXLC-3、GXLC-4、GXLC-5、GXLC-6、GXLC-7、GXLC-8、GXLC-9、GXLC-10、GXLC-11、GXLC-12、GXLC-13、GXLC-14、GXLC-15)和3株广西其他品种猪Mhp毒株(GX227、GX595、GX674)的P97基因R1区的多处碱基发生变异。通过推导氨基酸序列统计P97 R1区五氨基酸(AAKPV/E)重复数为9~18,平均值为12,TN重复数为1~4,结果发现广西陆川猪Mhp流行毒株变异使其毒力和黏附能力增强,再加上陆川猪呼吸道短小狭窄的特殊结构,更易导致广西陆川猪MPS的发生。

关键词: 广西陆川猪; 猪肺炎支原体; P97基因R1区; 克隆; 序列分析

Abstract: Through in-depth understanding of sequence characteristics, structure and genetic variation of adhesion factor P97 gene R1 region of Mycoplasma hyopneumoniae (Mhp) epidemic strains in Guangxi Luchuan pig, the assay was aimed to provide theoretical basis for mycoplasmal pneumonia of swine (MPS) integrated effective prevention and control measures of Guangxi local variety Luchuan pig, and lay a solid foundation for further study the characteristics of Mhp epidemic strains in Guangxi Luchuan pig.A pair of primers was designed to amplify P97 gene R1 region according to Mhp genome sequence, the MPS positive disease materials of Guangxi Luchuan pig from 2011 to 2014 as the object of study, after genomic DNA extraction and PCR amplification of P97 gene R1 region, the PCR products were sequenced.The base composition and deduced amino acid sequence of P97 gene R1 region of Mhp epidemic strains of Guangxi Luchuan pig were compared.The analysis of DNAStar showed that there were multiple base mutations in P97 gene R1 region of 15 strains of Guangxi Luchuan pig Mhp strains (GXLC-1, GXLC-2, GXLC-3, GXLC-4, GXLC-5, GXLC-6, GXLC-7, GXLC-8, GXLC-9, GXLC-10, GXLC-11, GXLC-12, GXLC-13, GXLC-14 and GXLC-15) and 3 strains of Guangxi other breeds of pigs Mhp strains (GX227, GX595 and GX674).The statistical repeat number of five amino acid (AAKPV/E) of P97 R1 region were 9 to 18, the average value was 12, TN repeats were 1 to 4.We found that Guangxi Luchuan pig Mhp strains mutation made its virulence and adhesion ability enhancement, and respiratory of Luchuan pig with the special structure of short and narrow, which could more easily lead to the occurrence of Guangxi Luchuan pig MPS.

Key words: Guangxi Luchuan pig; Mycoplasma hyopneumoniae; P97 gene R1 region; clone; sequence analysis

中图分类号:

黎珂钰, 赵武, 李斌, 秦毅斌, 卢冰霞, 梁家幸, 何颖, 周英宁, 韦英益. 广西陆川猪猪肺炎支原体P97基因R1区克隆及序列分析[J]. 《中国畜牧兽医》---唯一指定的官方网站, 2015, 42(9): 2270-2277.

LI Ke-yu, ZHAO Wu, LI Bin, QIN Yi-bin, LU Bing-xia, LIANG Jia-xing, HE Ying, ZHOU Ying-ning, WEI Ying-yi. Cloning and Sequence Analysis of P97 Gene R1 Region of Mycoplasma hyopneumoniae in Guangxi Luchuan Pig[J]. , 2015, 42(9): 2270-2277.

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广西陆川猪猪肺炎支原体P97基因R1区克隆及序列分析

Cloning and Sequence Analysis of P97 Gene R1 Region of Mycoplasma hyopneumoniae in Guangxi Luchuan Pig

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