SKLZΔspiC鸡白痢沙门菌减毒活疫苗候选株的遗传稳定性分析
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摘要
目的分析鸡白痢沙门菌减毒活疫苗候选株SKLZΔspiC的遗传稳定性。方法将疫苗株SKLZΔspiC与其亲体株S06004进行体外混合培养、体内共同感染,同时将重组质粒pMD20-spiC电转化导入疫苗株SKLZΔspiC,连续传至20代后,进行PCR及测序鉴定。结果在体外混合培养、体内混合感染条件下,疫苗株SKLZΔspiC均无法从野生株S06004获得靶基因spiC;疫苗株SKLZΔspiC无法从外源质粒获得靶基因spiC。结论疫苗株SKLZΔspiC具有良好的遗传稳定性。
Objective To analyze the genetic stability of candidate SKLZΔspiC strain for live attenuated Salmonella pullorum vaccine. Methods Candidate vaccine strain SKLZΔspiC and its parental strain S06004 were subjected to mixed culture in vitro and co-infection in vivo. Recombinant plasmid p MD20-spiC was transformed to vaccine strain SKLZΔspiC,and the obtained recombinant strain SKLZΔspiC(pMD20-spiC)was subcultured for 20 passages,and identified by PCR and sequencing. Results No target gene spiC was obtained from wild strain S06004 by the vaccine strain SKLZΔspiC in mixed infection in vitro and in vivo. Meanwhile,no spiC gene was obtained from foreign plasmids. Conclusion Candidate vaccine strain SKLZΔspiC showed high genetic stability.
Objective To analyze the genetic stability of candidate SKLZΔspiC strain for live attenuated Salmonella pullorum vaccine. Methods Candidate vaccine strain SKLZΔspiC and its parental strain S06004 were subjected to mixed culture in vitro and co-infection in vivo. Recombinant plasmid p MD20-spiC was transformed to vaccine strain SKLZΔspiC,and the obtained recombinant strain SKLZΔspiC(pMD20-spiC)was subcultured for 20 passages,and identified by PCR and sequencing. Results No target gene spiC was obtained from wild strain S06004 by the vaccine strain SKLZΔspiC in mixed infection in vitro and in vivo. Meanwhile,no spiC gene was obtained from foreign plasmids. Conclusion Candidate vaccine strain SKLZΔspiC showed high genetic stability.
引文
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[2] GENG S Z,LIU N N,JIAO X A,et al. Development and identification of spiC gene-deleted Salmonella pullorum mutant strainΔspiC[J]. Chin Vet Sci,2014,44(4):379-386.(in Chinese)耿士忠,刘男男,焦新安,等.鸡白痢沙门菌S06004ΔspiC突变株的构建与鉴定[J].中国兽医科学,2014,44(4):379-386.
[3] GENG S Z,LIU H,GUO R X,et al. Optimization of condition for storage of candidate bacterial strain of live attenuated Salmonella pullorum vaccine in frozen and genetic stability of stored strain[J]. Chin J Biologicals,2016,29(6):565-570.(in Chinese)耿士忠,刘欢,郭荣显,等.鸡白痢沙门菌减毒疫苗候选株冻存条件的优化及冻存菌株的遗传稳定性[J].中国生物制品学杂志,2016,29(6):565-570.
[4] GENG S Z,LIU H,JIAO X A,et al. Prokaryotic expression and application of SpiC protein of Salmonella pullorum[J].Chin J Cell Mol Immunol,2015,31(2):149-152,158.(in Chinese)耿士忠,刘欢,焦新安,等.鸡白痢沙门菌SpiC蛋白的原核表达及其应用[J].细胞与分子免疫学杂志,2015,31(2):149-152,158.
[5] SANAPALA S,RAHAV H,PATEL H,et al. Multiple antigens of Yersinia pestis delivered by live recombinant attenuated Salmonella vaccine strains elicit protective immunity against plague[J]. Vaccine,2016,34(21):2410-2416.
[6]耿士忠,郭荣显,焦新安,等.鸡白痢沙门菌研究进展[J].中国家禽,2014,36(5):34-37.
[7] PAN Z M,JIAO X A,HUANG J L,et al. Development of Salmonella gallinarum attenuated freeze dried vaccine[J]. Chin J Prev Vet Med,2001,23(6):444-447.(in Chinese)潘志明,焦新安,黄金林,等.鸡沙门氏菌弱毒冻干苗的研制[J].中国预防兽医学报,2001,23(6):444-447.
[8] WANG S,LI Y,SHI H,et al. Immune responses to recombinant pneumococcal PsaA antigen delivered by a live attenuated Salmonella vaccine[J]. Infect Immun,2015,78(7):3258-3271.
[9] LU G D,ZHANG K S,LIU Y J,et al. International standard of regulation and control for veterinary vaccine[J]. China Biotechnol,2011,31(5):138-141.(in Chinese)卢国栋,张克山,刘永杰,等.兽用疫苗管理与控制的国际标准[J].中国生物工程杂志,2011,31(5):138-141.
[10] PASCUAL D W,SUO Z,CAO L,et al. Attenuating gene expression(AGE)for vaccine development[J]. Virulence,2013,4(5):384-390.
[11] SVOBODOVJ,TMOVU E. Factors affecting microbial contamination of market eggs:a review[J]. Scientia Agr Bohemica,2015,45(4):226-237.
[12]徐仕忠.家禽沙门氏菌病及其疫苗控制[J].中国家禽,2006,28(17):48-50.
[13] ARNOLD M E,GOSLING R J,LA RAGIONE R M,et al.Estimation of the impact of vaccination on faecal shedding and organ and egg contamination for Salmonella enteritidis,Salmonella typhiumurium and monophasic Salmonella typhimurium[J]. Avian Pathol,2014,43(2):155-163.
[14] SIGGINS M K,O'SHAUGHNESSY C M,PRAVIN J. Differential timing of antibody-mediated phagocytosis and cell-free killing of invasive African Salmonella allows immune evasion[J]. Eur J Immunol,2014,44(4):1093-1098.
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