[1]嵇少泽,高云航*,勾长龙,等.牛多杀性巴氏杆菌znuA基因的原核表达及其免疫保护作用的研究[J].中国预防兽医学报,2019,(02):184-187.[doi:0.3969/j.issn.1008-0589.201807038]
 JI Shao-ze,GAO Yun-hang*,GOU Chang-long,et al.Prokaryotic expression of znuA of bovine Pasteurella multocidaand its immunoprotection in mice[J].Chinese journal of preventive veterinary medicine,2019,(02):184-187.[doi:0.3969/j.issn.1008-0589.201807038]
点击复制

牛多杀性巴氏杆菌znuA基因的原核表达及其免疫保护作用的研究()
分享到:

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

卷:
期数:
2019年02
页码:
184-187
栏目:
免疫学
出版日期:
2019-02-15

文章信息/Info

Title:
Prokaryotic expression of znuA of bovine Pasteurella multocida
and its immunoprotection in mice
文章编号:
1008-0589(2019)02-0184-04
作者:
 

嵇少泽高云航*勾长龙张喜庆田佳琪李 晨王思月

 (吉林农业大学 动物科学技术学院,吉林 长春 130118)
Author(s):
 

JI Shao-ze GAO Yun-hang* GOU Chang-long ZHANG xi-qing TIAN Jia-qi LI Chen WANG Si-yue

 (Department of Preventive Veterinary Medicine, Jilin Agricultural University, Changchun 130118, China)

 

关键词:
多杀性巴氏杆菌znuA保护性免疫
Keywords:
 Pasteurella multocida  znuA  protective immunity
分类号:
S852.61
DOI:
0.3969/j.issn.1008-0589.201807038
文献标志码:
A
摘要:
为研究牛多杀性巴氏杆菌(P.multocida)高亲和力锌吸收蛋白znuA的免疫学活性及其免疫保护作用,本研究利用 PCR方法扩增了P.multocida znuA基因,构建表达载体 pET-28a-znuA,将其转化E.coli BL21 后经诱导表达。表达产物经SDS-PAGE和western blot分析显示,重组蛋白约 40 ku;以重组znuA蛋白免疫小鼠后用P.multocida菌株Y-1攻毒,结果显示重组znuA蛋白对免疫组小鼠保护率为 60 %,表明其具有免疫保护作用。本研究首次在原核系统中表达了P.multocida znuA蛋白,且验证了其免疫保护力,为深入探究znuA基因在P.mul- tocida致病过程中的作用及其亚单位疫苗的开发奠定了基础。
  
Abstract:
To study the immunoprotection of high-affinity zinc uptake system protein znuA against in mice. The target gene,znuA, was amplified by PCR from Pasteurella multocida of Bovine and cloned into the pET-28a vector. The constructed recombinant plasmid pET-28a-znuA was transformed to E.coli BL21(DE3)for expression under induction of IPTG. SDS-PAGE and western blot analysis showed that the expressed recombinant znuA was about 40ku. The mice were immunized intramuscularly with purified recombinant znuA, and challenged with the Pasteurella multocida Y-1, result showed that the recombinant znuA provided a 60% protection for the immunized mice. All results indicated that the target protein can be over expressed in E.coli BL21, which would induced protective immune response in mice, indicating its potential as an antigen of Pasteurella multocida protein vaccine.

参考文献/References:

[1] Harper M, Boyce J D, Adler B. Pasteurella multocida pathogenesis: 125 years after Pasteur [J]. FEMS Microbiol Lett, 2006, 265(1): 1-10.
[2]Dziva F, Muhairwa A P, Bisgaard M, et al. Diagnostic and typing options for investigating diseases associated with Pasteurella multocida [J]. Vet Microbiol, 2008, 128(1-2): 1-22.
[3]Banerjee S, Wei B, Bhattacharyyapakrasi M, et al. Structural determinants of metal specificity in the zinc transport protein ZnuA from synechocystis 6803 [J]. J MBio, 2003, 333(5): 1061-1069.
[4]Yang Xing-hong, Becker T, Walters N, et al. Deletion of znuA virulence factor attenuates Brucella abortus and confers protection against wild-type challenge [J]. Infect Immun, 2006, 74(7): 3874-3879.
[5]Kim S, Watanabe K, Shirahata T, et al. Zinc uptake system (znuA locus) of Brucella abortus is essential for intracellular survival and virulence in mice [J]. J Vet Med Sci, 2004, 66(9): 1059-1063.
[6]Lewis D A, Klesneytait J, Lumbley S R, et al. Identification of the znuA-encoded periplasmic zinc transport protein of Haemo- philus ducreyi [J]. Infect Immun, 1999, 67(10): 5060-5068.
[7]Ammendola S, Pasquali P, Pistoia C, et al. High-Affinity Zn2+ uptake system ZnuABC is required for bacterial Zinc homeostasis in intracellular environments and contributes to the virulence of Salmonella enterica [J]. Infect Immun, 2007, 75(12): 5867- 5876.
[8]Chen C Y, Morse S A. Identification and characterization of a high-affinity zinc uptake system in Neisseria gonorrhoeae [J]. FEMS Microbiol Lett, 2001, 202(1): 67-71.
[9]李洪涛,张新涛,刘明,等. A型禽流感病毒不同拷贝数基质蛋白胞外区基因的原核表达及免疫保护力研究[J]. 中国预防兽医学报,2010,32(10):795-799.
[10]Tang Jia-qi, Wang Chang-jun, Feng You-jun, et al. Streptococcal toxic shock syndrome caused by Streptococcus suis serotype 2 [J]. PLoS Med, 2006, 3(5): e151.
[11]Janulczyk R, Ricci S, Bj?觟rck L. MtsABC is important for manganese and iron transport, oxidative stress resistance, and virulence of Streptococcus pyogenes [J]. Infect Immun, 2003, 71(5): 2656-2564.
[12]Garmory H S, Titball R W. ATP-binding cassette transporters are targets for the development of antibacterial vaccines and therapies [J]. Infect Immun, 2004, 72(12): 6757-6763.
[13]Staats J J, Feder I, Okwumabua O, et al. Streptococcus suis: past and present [J]. Vet Res Commun, 1997, 21(6): 381-407.
[14]Bartelink A K, Van K E. Streptococcus suis as threat to pig- farmers and abattoir workers [J]. Lancet, 1995, 346(8991-8992): 1707.
[15]Gottschalk M, Segura M, Xu J. Streptococcus suis infections in humans: the Chinese experience and the situation in North America [J]. Ani Heal Res Rev, 2007, 8(1): 29-45.
[16]Olsen W A, Follmann F, Erneholm K, et al. Protection against chlamydia trachomatis infection and upper genital tract pathological changes by vaccine-promoted neutralizing antibodies directed to the VD4 of the major outer membrane protein [J]. J Infect Dis, 2015, 212(6): 978.
[17]刘丽娜,张洁,周静,等. 金黄色葡萄球菌疫苗候选分子StbA主要抗原表位区的表达及重组蛋白抗血清的制备[J]. 中国兽医科学,2015,(4):375-379.
[18]韩明月,潘秀珍,邵珠卿,等. 2型猪链球菌中国强毒株znuA基因的原核表达及免疫学活性分析[J]. 免疫学杂志,2010,(3):220-223.
                                (本文编辑:彭永刚)

相似文献/References:

[1]姜志刚,马文戈,于 力*.牛源A型多杀性巴氏杆菌培养特性和免疫原性的研究[J].中国预防兽医学报,2010,(03):183.
 JIANG Zhi-gang,MA Wen-ge,YU Li*.Cultural characterization and immunogenicity of bovine capsular serotype A Pasteurella mutocida[J].Chinese journal of preventive veterinary medicine,2010,(02):183.
[2]郭东春,孙 yan,刘家森,等.鸭多杀性巴氏杆菌外膜蛋白H基因的表达及抗原性鉴定[J].中国预防兽医学报,2010,(07):574.
 GUO Dong-chun,SUN Yan,LIU Jia-sen,et al.Expression and antigenicity analyasis of the Omp H Gene of Pasteurella multocida[J].Chinese journal of preventive veterinary medicine,2010,(02):574.
[3]马文戈,于 力*.牛源荚膜血清A型多杀性巴氏杆菌的分离鉴定[J].中国预防兽医学报,2008,(10):747.
 MA Wen-ge,YU Li*.Isolation and identification of bovine capsular serotype A Pasteurella mutocida[J].Chinese journal of preventive veterinary medicine,2008,(02):747.
[4]李永明,罗阿东,徐景峨,等.猪源多杀性巴氏杆菌的生物学鉴定与荚膜PCR分型[J].中国预防兽医学报,2007,(07):506.
 LI Yong-ming,LUO A-dong,XU Jing-e,et al.PCR identification and typing of Pasteurella multocida originated from pigs[J].Chinese journal of preventive veterinary medicine,2007,(02):506.
[5]孙翠平,王金良,李书光,等.雏鸭源多杀性巴氏杆菌的分离鉴定和荚膜PCR分型[J].中国预防兽医学报,2011,(07):521.
 SUN Cui-ping,WANG Jin-liang,LI Shu-guang,et al.Isolation and identification of a Pasteurella mutocida from ducklings[J].Chinese journal of preventive veterinary medicine,2011,(02):521.
[6]田 亮,康立超*,赵文娟,等.牦牛源多杀性巴氏杆菌血清型鉴定及ompH基因序列分析[J].中国预防兽医学报,2012,(02):152.[doi:10.3969/j.issn.1008-0589.2012.02.18]
 TIAN Liang,KANG Li-chao *,ZHAO Wen-juan,et al.Identified the serotype and sequences analysis of the yak ompH gene of Pasteurellosis multocida[J].Chinese journal of preventive veterinary medicine,2012,(02):152.[doi:10.3969/j.issn.1008-0589.2012.02.18]
[7]季权安,刘 燕,庞安娜,等.兔多杀性巴氏杆菌16S rRNA PCR检测方法的建立[J].中国预防兽医学报,2012,(06):468.[doi:doi: 10.3969/j.issn.1008-0589.2012.06.12]
 JI Quan-an,LIU Yan,PANG An-na,et al.Establishment of PCR assay based on 16S rRNA forthe diagnosis of Pasteurella multocida infection in rabbits[J].Chinese journal of preventive veterinary medicine,2012,(02):468.[doi:doi: 10.3969/j.issn.1008-0589.2012.06.12]
[8]周 伟,李能章,魏学良,等.雁鸭源荚膜血清A型多杀性巴氏杆菌的分离鉴定[J].中国预防兽医学报,2012,(06):498.[doi:doi: 10.3969/j.issn.1008-0589.2012.06.20]
 ZHOU Wei,LI Neng-zhang,WEI Xue-liang,et al.Isolation and identification of Anatidae capsular serotype A Pasteurella mutocida[J].Chinese journal of preventive veterinary medicine,2012,(02):498.[doi:doi: 10.3969/j.issn.1008-0589.2012.06.20]
[9]卢 艳,郭东春,张爱芹,等.抗鸭多杀性巴氏杆菌外膜蛋白H单克隆抗体的制备及鉴定[J].中国预防兽医学报,2012,(10):833.[doi:doi: 10.3969/j.issn.1008-0589.2012.10.19]
 LU Yan,GUO Dong-chun,ZHANG Ai-qin,et al.Preparation and identification of the monoclonal antibody against outer membrane proteins H of Pasteurella multocida[J].Chinese journal of preventive veterinary medicine,2012,(02):833.[doi:doi: 10.3969/j.issn.1008-0589.2012.10.19]
[10]王淑亚,郭东春,李 影,等.多杀性巴氏杆菌Lon蛋白酶的原核表达及ATPase活性的检测[J].中国预防兽医学报,2014,(07):530.[doi:10.3969/j.issn.1008-0589.2014.07.07]
 WANG Shu-ya,GUO Dong-chun,LI Ying,et al.Prokaryotic expression and detection of the ATPase activityof Lon protease from Pasteurella multocida[J].Chinese journal of preventive veterinary medicine,2014,(02):530.[doi:10.3969/j.issn.1008-0589.2014.07.07]

更新日期/Last Update: 2019-03-07