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| 表达T~+Pm保护性抗原的重组猪霍乱沙门氏菌C500株的构建及其生物学特性 | |
| 引用本文： | 赵战勤,王臣,丁轲,李晓峰,吴斌,卢顺,张春杰,程相朝. 表达T~+Pm保护性抗原的重组猪霍乱沙门氏菌C500株的构建及其生物学特性[J]. 微生物学报, 2010, 50(1): 91-97 |
| 作者姓名： | 赵战勤 王臣 丁轲 李晓峰 吴斌 卢顺 张春杰 程相朝 |
| 作者单位： | 1. 河南科技大学动物科技学院,动物病原微生物学实验室,洛阳471003;华中农业大学动物医学院,农业微生物学国家重点实验室,武汉430070 2. 河南科技大学动物科技学院,动物病原微生物学实验室,洛阳471003 3. 河南农业大学牧医工程学院,郑州,450002 4. 华中农业大学动物医学院,农业微生物学国家重点实验室,武汉430070 |
| 基金项目： | 河南科技大学博士基金项目（090001337）；国家“863计划”(2006AA10A206) |
| 摘 要： | 【目的】本研究利用Asd+平衡致死系统构建表达巴氏杆菌毒素(Pasteurella multocida toxin,PMT)的重组猪霍乱沙门氏菌株,并对重组菌株的生物学特性进行比较研究。【方法和结果】通过基因克隆的方法构建表达PMT的重组质粒pYA-PmtC,再将其电转化减毒猪霍乱沙门氏菌C500的asd基因缺失株C501,构建口服活疫苗菌株C501(pYA-PmtC)。研究结果表明重组菌株C501(pYA-PmtC)的生化特性、血清型和生长速度与亲本菌株C500一致;在没有选择压力的条件下,C501(pYA-PmtC)能够稳定遗传重组质粒及其外源基因片段,并能稳定、高效、分泌性表达30.5kDa的外源保护性抗原rPmtC。C501(pYA-PmtC)腹腔感染BALB/c小鼠的LD50为8.5×106CFU,毒力稍低于C500(LD50为4.4×106CFU);口服接种C501(pYA-PmtC)和C500的所有仔猪未见任何发病症状,两者没有显著差别。【结论】本研究利用Asd+平衡致死系统的原理构建表达T+Pm保护性抗原重组猪霍乱沙门氏菌弱毒菌株C501(pYA-PmtC),为进一步开发猪萎缩性鼻炎-副伤寒的双价基因工程疫苗奠定基础。 |
| 关 键 词： | 关键词：猪霍乱沙门氏菌C500株，产毒巴氏杆菌，Asd+平衡致死系统，重组疫苗 |
| 收稿时间： | 2009-07-03 |
| 修稿时间： | 2009-09-09 |
Construction and characterization of recombinant Salmonella enterica serovar choleraesuis vaccine strain expressing heterologous antigen of Pasteurella multocida toxin |
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| Zhanqin Zhao,Chen Wang,Ke Ding,Xiaofeng Li,Bin Wu,Shun Lu,Chunjie Zhang and Xiangchao Cheng. Construction and characterization of recombinant Salmonella enterica serovar choleraesuis vaccine strain expressing heterologous antigen of Pasteurella multocida toxin[J]. Acta microbiologica Sinica, 2010, 50(1): 91-97 | |
| Authors: | Zhanqin Zhao Chen Wang Ke Ding Xiaofeng Li Bin Wu Shun Lu Chunjie Zhang Xiangchao Cheng |
| Affiliation: | Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China;Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China;Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China;College of Animal Husbandry and Veterinary Science, Henan Agriculture University, Zhengzhou 450002, China;The State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China;The State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China;Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China;Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China |
| Abstract: | Abstract: [Objective] This study was aimed to construct and characterize Salmonella vaccine strain C500 expressing the recombinant Pasteurella multocida toxin (PMT) antigen by the Asd+ balanced-lethal host-vector system. [Methods and results] The DNA fragment encoding C terminal of PMT was cloned downstream from the beta-lactamase signal sequence in the multicopy Asd+ pYA3493 vector to create pYA-PmtC. Fermentation patterns, serotype, and mean generation time of the vaccine strain C500 harboring pYA-PmtC (named with C501(pYA-PmtC)) were identical to those of the parent strain C500. The recombinant pYA-PmtC plasmid was very stable in C501(pYA-F1P2), which expressed secretorily a large amount of the recombinant PMT antigen (named with rPmtC). The virulence of C501(pYA-PmtC) with LD50 of 8.5×106 CFU was a little lower than C500 with LD50 of 4.4×106 CFU based on the method of Reed and Muench. All piglets inoculated with C501(pYA-PmtC) or C500 survived, and had no signs of disease during the entire experimental period. No significant differences were found between these two groups. [Conclusion] The recombinant vaccine strain C501(pYA-PmtC) had a series of biological characteristics silimar to the parent vaccine strain C500. It is likely that C501(pYA-PmtC) could be adapted to develop multivalent recombinant Salmonella vaccine against both infections with S. enterica serovar Choleraesuis and toxigenic P. multocida. |
| Keywords: | Keywords: Salmonella enterica Serovar Choleraesuis C500 strain toxigenic Pasteurella multocida Asd+ balanced-lethal host-vector system recombinant vaccine |
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