Ⅰa型牛源无乳链球菌LZQ07006分离株BibA基因片段的分子特征分析

中国畜牧兽医

Ⅰa型牛源无乳链球菌LZQ07006分离株BibA基因片段的分子特征分析

常瑞祥，王旭荣，王磊，张景艳，秦哲，孔晓军，王孝武，李建喜，杨志强，王学智

（中国农业科学院兰州畜牧与兽药研究所，甘肃省中兽药工程技术研究中心，甘肃兰州 730050）

修回日期:2014-03-04 出版日期:2014-05-20 发布日期:2014-06-25
通讯作者: 杨志强（1957—），男，研究员，主要从事奶牛病研究。E-mail：zhiqyang20062006@163.com；Tel：0931-2115258 王学智（1969—），男，博士，副研究员，主要从事兽医临床诊断学研究。E-mail：wangxuezhi@caas.cn；Tel：0931-2115196
作者简介:常瑞祥（1987—），男，河南人，硕士生，从事奶牛乳房炎细菌研究。
基金资助:
国家奶牛产业技术体系科学家岗位项目（CARS37）；甘肃省农业生物技术研究与应用开发项目（GNSW-2013-28）；公益性行业（农业）科研专项（201303040-01）。

Molecular Characterization Analysis of BibA Gene in the Isolate LZQ07006 of Serotype Ⅰa Streptococcus agalactiae from Bovine

CHANG Rui-xiang, WANG Xu-rong, WANG Lei, ZHANG Jing-yan, QIN Zhe, KONG Xiao-jun, WANG Xiao-wu, LI Jian-xi, YANG Zhi-qiang, WANG Xue-zhi

(Engineering & Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS, Lanzhou 730050, China)

Revised:2014-03-04 Online:2014-05-20 Published:2014-06-25

摘要/Abstract

摘要： 试验旨在对Ⅰa型牛源无乳链球菌BibA基因及其编码的蛋白进行分子特征分析，为进一步研究BibA蛋白的表达及免疫原性提供理论依据。采用PCR技术扩增牛源无乳链球菌LZQ07006分离株BibA基因片段，并将其克隆入pMD20-T载体后测序，采用生物学软件对BibA基因及其推导的蛋白进行生物信息学分析。结果显示，LZQ07006菌株BibA基因片段长度为1185 bp，未出现基因缺失，编码395个氨基酸，与GenBank中已发表的不同血清型的无乳链球菌核苷酸同源性和推导的氨基酸同源性分别为45.0%～99.8%和12.3%～99.5%；BibA基因片段是一个稳定的分泌性外膜蛋白，亲水性较好，存在1—32位氨基酸的信号肽，剪切位点在第32—33位氨基酸之间的蛋白片段上。因此，牛源无乳链球菌BibA蛋白可作为奶牛乳房炎诊断和预防的应用性候选蛋白，但需深入研究。

关键词: font-size: 10.5pt; mso-bidi-font-size: 12.0pt; mso-font-kerning: 1.0pt; mso-bidi-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA">Ⅰa型；无乳链球菌；BibAfont-size: 10.5pt; mso-bidi-font-size: 12.0pt; mso-font-kerning: 1.0pt; mso-bidi-font-family: 宋体; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA">基因；克隆；序列分析

Abstract: This study was aimed at analyzing the molecular characteristics of BibA gene and its expression protein, providing theoretical basis for the further study on the expression and immunogenicity of BibA protein. The BibA gene of the isolate LZQ07006 of serotype Ⅰa Streptococcus agalactiae from bovine was amplified using the PCR method, cloned into pMD20-T vector and then sequenced. The molecular characteristics of BibA gene and its expression protein were analyzed using a variety of biological software. The results showed that BibA gene of LZQ07006 was 1185 bp and did not appear the gene deletion, and encoded 395 amino acids. The nucleotide sequence homology of BibA gene derived from LZQ07006 was 45.0% to 99.8% and the amino acid homology was 12.3% to 99.5% among the corresponding sequences from different serotypes Streptococcus agalactiae published in GenBank. The derived amino acid showed that the BibA protein was a stable outer membrane protein, which was secretion protein and hydrophilic highly. The polypeptide contained a signal peptide consisting of 32 amino acids, with cleavage site between 32th and 33th amino acids. In conclusion, it was necessary to study the BibA protein in bovine Streptococcus agalactiae for application in diagnosis or prevention of dairy cow mastitis.

Key words: font-size: 10.5pt; mso-bidi-font-size: 12.0pt; mso-font-kerning: 1.0pt; mso-bidi-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA" lang="EN-US">serotype font-size: 10.5pt; mso-bidi-font-size: 12.0pt; mso-font-kerning: 1.0pt; mso-bidi-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA">Ⅰa; Streptococcus agalactiae; BibA gene; clone; sequence analysis

常瑞祥，王旭荣，王磊，张景艳，秦哲，孔晓军，王孝武，李建喜，杨志强，王学智. Ⅰa型牛源无乳链球菌LZQ07006分离株BibA基因片段的分子特征分析[J]. 中国畜牧兽医.

CHANG Rui-xiang, WANG Xu-rong, WANG Lei, ZHANG Jing-yan, QIN Zhe, KONG Xiao-jun, WANG Xiao-wu, LI Jian-xi, YANG Zhi-qiang, WANG Xue-zhi. Molecular Characterization Analysis of BibA Gene in the Isolate LZQ07006 of Serotype Ⅰa Streptococcus agalactiae from Bovine[J]. .

参考文献

1 王旭荣，张世栋，杨峰，等. Ⅰa型和Ⅱ型牛源无乳链球菌sip基因的遗传进化分析［J］. 中国兽医科学，2012，42(9)：888～894.
2 吴金花. 通辽地区奶牛乳房炎主要病原菌分离及药物敏感性试验［J］. 黑龙江畜牧兽医，2007(3)：77～78.
3 李宏胜，郁杰，李新圃，等. 奶牛乳房炎类型与病原菌感染之间相关性的研究［J］. 动物医学进展，2004，25(6)：80～84.
4 李宏胜，郁杰，罗金印，等．牛源性无乳链球菌血清型分布及抗生素耐药性检测［J］. 中国畜牧兽医，2012，39(1)：164～166.
5 常瑞祥，王旭荣，杨志强，等. 无乳链球菌黏附因子研究进展［J］. 动物医学进展，2014，35(2)：101～104.
6 Abera M, Habte T, Aragaw K, et al. Major cause of mastitis and associated risk factors in smallholder dairy farms in and around Hawassa, Southern Ethiopia［J］. Trop Anim Health Prod, 2012, 44(6): 1175～1179.
7 Dogan B, Schukken Y H, Santisteban C, et al. Distribution of serotypes and antimicrobial resistance genes among Streptococcus agalactiae isolates from bovine and human hosts［J］. J Clin Microbiol, 2005, 43(12): 5899～5906.
8 Edmond K M, Kortsalioudaki C, Scott S, et al. Group B streptococcal disease in infants aged younger than 3 months: Systematic review and meta-analysis［J］. Lancet, 2012, 379(9815): 547～556.
9 Edwards M S, Baker C J. Group B Streptococcal infections in elderly adults［J］. Clin Infect Dis, 2005, 41(6): 839～847.
10 Heath P T. An update on vaccination against group B Streptococcus［J］. Expert Rev Vaccines, 2011, 10(5): 685～694.
11 Oliveira I C, de Mattos M C, Pinto T A, et al. Genetic relatedness between group B streptococci originating from bovine mastitis and a human group B Streptococcus type V cluster displaying an identical pulsed-field gel electrophoresis pattern［J］. Clin Microbiol Infect, 2006, 12(9): 887～893.
12 Phares C R, Lynfield R, Farley M M, et al. Epidemiology of invasive group B streptococcal disease in the United States, 1999—2005［J］. Jama, 2008, 299(17): 2056～2065.
13 Richards V P, Choi S C, Bitar P D, et al. Transcriptomic and genomic evidence for Streptococcus agalactiae adaptation to the bovine environment［J］. BMC Genomics, 2013, 14: 920.
14 Santi I, Scarselli M, Mariani M, et al. BibA: A novel immunogenic bacterial adhesin contributing to group B Streptococcus survival in human blood［J］. Mol Microbiol, 2007, 63(3): 754～767.
15 Santi I, Maione D, Galeotti C L, et al. BibA induces opsonizing antibodies conferring in vivo protection against group B Streptococcus［J］. J Infect Dis, 2009, 200(4): 564～570.
16 Schrag S J, Zywicki S, Farley M M, et al. Group B streptococcal disease in the era of intrapartum antibiotic prophylaxis［J］. N Engl J Med, 2000, 342(1): 15～20.
17 Sentitula, Yadav B R, Kumar R. Incidence of staphylococci and Streptococcus agalactiea during winter in mastitis milk of Sahiwal cow and murrah Buffaloes［J］. Indian J Microbil, 2012, 52(2): 153～159.
18 Skoff T H, Farley M M, Petit S, et al. Increasing burden of invasive group B streptococcal disease in nonpregnant adults, 1990—2007［J］. Clin Infect Dis, 2009, 49(1): 85～92.
19 Tazi A, Morand P C, Réglier-Poupet H, et al. Invasive group B streptococcal infections in adults, France (2007—2010)［J］. Clin Microbiol Infect, 2011, 17(10): 1587～1589.
20 Tettelin H, Masignani V, Cieslewicz M J, et al. Genome analysis of multiple pathogenic isolates of Streptococcus agalactiae: Implications for the microbial “pan-genome”［J］. Proc Natl Acad Sci USA, 2005, 102(39): 13950～13955.

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