[1]汤 君,文心田,曹三杰*,等.猪传染性胸膜肺炎放线杆菌转铁结合蛋白B基因的克隆与表达[J].中国预防兽医学报,2007,(12):943-945.
 TANG Jun,WEN Xin-tian,CAO San-jie*,et al.Cloning and expression of the tbpB gene of Actinobacillus pleuropneumoniae[J].Chinese journal of preventive veterinary medicine,2007,(12):943-945.
点击复制

猪传染性胸膜肺炎放线杆菌转铁结合蛋白B基因的克隆与表达
分享到:

《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2007年12期
页码:
943-945
栏目:
病原生物学
出版日期:
2007-12-01

文章信息/Info

Title:
Cloning and expression of the tbpB gene of Actinobacillus pleuropneumoniae
文章编号:
1008-0589(2007)12-0943-03
作者:
汤 君文心田曹三杰*黄小波杨 利
四川农业大学动物科技学院 动物传染病与基因芯片实验室/动物疫病与人类健康四川省重点实验室,四川 雅安 625014
Author(s):
TANG Jun WEN Xin-tian CAO San-jie* HUANG Xiao-bo YANG Li
Laboratory of Animal Infectious Disease and Microarray, Key Laboratory of Animal Disease and Human Health of Sichuan Province, College of Animal Science and Veterinary Medicine, Sichuan Agricultural University, Ya’an 625014, China
关键词:
胸膜肺炎放线杆菌转铁结合蛋白B克隆表达
Keywords:
pig Actinobacillus pleuropneumoniae tbpB cloning expression
分类号:
S852.619;Q786
文献标志码:
A
摘要:
参照猪传染性胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)血清1型菌株,设计1对特异性引物,PCR扩增转铁结合蛋白B(tbpB)全基因,克隆到pMD19-T Simple载体中,经测序比较,与参考序列的核苷酸同源性达99.72 %。试验将tbpB基因定向克隆到pET-32a(+)中,转化BL21(DE3),经诱导后,SDS-PAGE结果显示转铁结合蛋白B得到表达,Western blot检测呈阳性。
Abstract:
The tbpB gene from serotype 1 strain of Actinobacillus pleuropneumoniae was amplified by PCR. The amplified DNA fragment was cloned into pMD19-T simple, and sequenced. The result showed that the gene was 1 812 bp in length and shared 99.72 % sequence identity with that of standard strains. The gene was subcloned into in pET-32a (+) and successfully expressed in Escherichia coli BL21 (DE3) after induced, which was confirmed by Western blot.

参考文献/References:

[1] B E Straw S D,Allaire W L,Mengeling D J,et al. 猪病学[M]. 第8版. 北京:中国农业大学出版社,2000.
[2] 张立昌. 猪传染性胸膜肺炎研究进展[J]. 养猪,2001,(1):40-42.
[3] Tascon R I, Vazquez-Boland J A, Gutierrez-Martin C B, et al. Virulence factors of the swine pathogen Actinobacillus pleuropn- eumoniae [J]. Microbiologia, 1996, 12: 171-184.
[4] Deneer H G, Potter A A. Effect of iron restriction on the outer membrane proteins of Actinobacillus (Haemophilus) pleuropneumoniae [J]. Infect Immun, 1989, 57: 798-804.
[5] Baltes N, Hennig-Pauka I, Gerlach G F. Both transferrin binding proteins are virulence factors in Actinobacillus pleuropneumoniae serotype 7 infection [J]. FEMS Microbiol Lett, 2002, 209(2): 283-287.
[6] Kim T, Lee J. Cloning and expression of genes encoding transferrin-binding protein A and B from Actinobacillus pleuropneumoniae serotype 5 [J]. Protein Expr Purif, 2006, 45(1): 235-240.
[7] J萨姆布鲁克,E F弗里奇,T曼尼阿蒂斯. 分子克隆实验指南[M]. 第2版. 金冬雁,黎孟枫,侯云德,等译. 北京:科学出版社,1992.
[8] Bosse J T, Janson H, Sheehan B J, et al. Actinobacillus pleuropneumoniae: pathobiology and pathogenesis of infection [J]. Microbes Infect, 2002, 4: 225-235.
[9] Rossi-Campos A, Anderson C, Gerlach G F, et al. Immunization of pigs against Actinobacillus pleuropneumoniae with two recombinant protein preparations [J]. Vaccine, 1992, 10(8): 512-518.

相似文献/References:

[1]蒋媛媛,郭军庆,颜焰,等.抗猪lgG单克隆抗体的制备与应用[J].中国预防兽医学报,2008,(09):727.
[2]徐公义,王海丽,葛长城,等.鲁西地区猪胸膜肺炎放线杆菌血清型多重PCR研究 [J].中国预防兽医学报,2009,(06):454.
 XU Gong-yi,WANG Hai-li,GE Chang-cheng,et al.Epidemiological investigation of Actinobacillus Pleuropneaumoniae serotype in western Shandong by multiplex PCR [J].Chinese journal of preventive veterinary medicine,2009,(12):454.
[3]杨 鹏,雷连成*,谢 芳,等.痤疮丙酸杆菌对小鼠抗胸膜肺炎放线杆菌感染的研究[J].中国预防兽医学报,2009,(08):596.
 YANG Peng,LEI Lian-cheng*,XIE Fang,et al.Immune activity of Propionibacterium acnes against Actinobacillus pleuropneumoniae infection in mice [J].Chinese journal of preventive veterinary medicine,2009,(12):596.
[4]高 明,魏 华*.重组猪IFN-α原核表达及其活性分析 [J].中国预防兽医学报,2009,(11):902.
 GAO Ming,WEI Hua*.Expression of recombinant porcine alpha-interferon in E.coli and the anti-virus activity analysis[J].Chinese journal of preventive veterinary medicine,2009,(12):902.
[5]施开创,李焕荣,杨汉春*,等.猪IL-4、IL-6和IL-10 SYBR Green I real-time PCR检测方法的建立及应用[J].中国预防兽医学报,2010,(02):102.
 SHI Kai-chuang,LI Huan-rong,YANG Han-chun*,et al.Development and application of a SYBR Green I-based real-time PCR assay for detection of porcine IL-4, IL-6 and IL-10[J].Chinese journal of preventive veterinary medicine,2010,(12):102.
[6]王贵平,刘军发,陈剑锋,等.胸膜肺炎放线杆菌荧光抗体检测方法的建立[J].中国预防兽医学报,2008,(12):949.
 WANG Gui-ping,LIU Jun-fa,CHEN Jian-feng,et al.Development of species-specific immunofluorescence assay to detect Actinobacillus pleuopneumoniae[J].Chinese journal of preventive veterinary medicine,2008,(12):949.
[7]张莉莉,白 娟,王先炜,等.猪Toll样受体3、7和8基因的克隆与序列分析[J].中国预防兽医学报,2010,(09):727.
 ZHANG Li-li,BAI Juan,WANG Xian-wei,et al.Cloning and sequencing analysis of porcine Toll-like receptor 3, 7 and 8 genes[J].Chinese journal of preventive veterinary medicine,2010,(12):727.
[8]谢 芳,韩文瑜,杜崇涛,等.胸膜肺炎放线杆菌血清1型与5型基因组DNA差异基因的筛选与鉴定[J].中国预防兽医学报,2010,(10):757.[doi:10.3969/j.issn.1008-0589.2010.10.04]
 XIE Fang,HAN Wen-yu,DU Chong-tao,et al.Screening and identification of the differential genomic DNA between Actinobacillus pleuropneumoniae serotype 1 and serotype 5[J].Chinese journal of preventive veterinary medicine,2010,(12):757.[doi:10.3969/j.issn.1008-0589.2010.10.04]
[9]王 芳,何孔旺*,邱索平,等.猪传染性胸膜肺炎放线杆菌毒素I基因的克隆、序列分析及表达[J].中国预防兽医学报,2006,(02):121.
 WANG Fang,HE Kong-wang*,QIU Suo-ping,et al.Cloning,sequencing and expression of the apxⅠgene of Actinobacillus pleuropneumonia[J].Chinese journal of preventive veterinary medicine,2006,(12):121.
[10]王 新*,黄 山,周 婷,等.猪源耐甲氧西林金黄色葡萄球菌的耐药性及其SCCmec基因分型研究[J].中国预防兽医学报,2010,(12):975.[doi:10.3969/j.issn.1008-0589.2010.12.16]
 WANG Xin*,HUANG Shan,ZHOU Ting,et al.Antimicrobial susceptibility and SCCmec genetyping of methicillin-resistant Staphylococcus aureus isolates from swine[J].Chinese journal of preventive veterinary medicine,2010,(12):975.[doi:10.3969/j.issn.1008-0589.2010.12.16]

备注/Memo

备注/Memo:
收稿日期:2007-02-18
基金项目:四川省科技攻关重大项目(05NG002-012和05NG020-016)
作者简介:汤 君(1984~),男,重庆人,硕士研究生,主要从事预防兽医学及分子生物学研究.
*通讯作者:E-mail:csanjie@sicau.edu.cn
更新日期/Last Update: 2011-01-06