| 表达空肠弯曲菌CfrA蛋白重组乳酸乳球菌的构建及其免疫效果北大核心CSCD |
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| 引用本文： | 王传文,周宏专,郭芳芳,陈玉霞,杨兵,苏霞,朱瑞良,徐福洲. 表达空肠弯曲菌CfrA蛋白重组乳酸乳球菌的构建及其免疫效果北大核心CSCD[J]. 微生物学报, 2016, 56(8): 1326-1334 |
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| 作者姓名： | 王传文 周宏专 郭芳芳 陈玉霞 杨兵 苏霞 朱瑞良 徐福洲 |
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| 作者单位： | 山东农业大学动物科技学院, 山东省动物生物技术与疫病防治重点实验室, 山东 泰安 271018;北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097,山东农业大学动物科技学院, 山东省动物生物技术与疫病防治重点实验室, 山东 泰安 271018,北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097 |
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| 基金项目： | 国家自然科学基金（31172344）；北京市农林科学院科技创新能力建设专项（KJCX20150703） |
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| 摘 要： | 【目的】本试验将空肠弯曲菌肠菌素受体蛋白CfrA编码基因导入食品级乳酸乳球菌表达系统,然后将重组乳酸乳球菌口服免疫鸡,降低空肠弯曲菌在鸡肠道中的定殖。【方法】利用PCR分别扩增空肠弯曲菌cfrA全基因及其N端片段,插入食品级表达载体pNZ8149多克隆位点并转化乳酸乳球菌NZ3900,通过Western blot鉴定重组菌株CfrA蛋白表达情况,同时通过筛选nisin浓度、温度、时间等诱导条件优化重组蛋白表达水平;进而将重组乳酸乳球菌经口服免疫SPF鸡,免疫后分别测定乳酸乳球菌自鸡体内的排出情况、以及诱导CfrA血清抗体和粘膜抗体水平,最后将空肠弯曲菌口服攻毒免疫后的鸡,通过测定鸡泄殖腔棉拭子中空肠弯曲菌的数目来判定口服免疫效果。【结果】Western blot检测显示CfrA全基因及其N端片段均可在重组乳酸乳球菌胞内可溶性表达,不分泌,筛选的最佳诱导表达条件为nisin浓度25 ng/mL、温度37°C、时间1 h。口服乳酸乳球菌10 d内自鸡体完全排空;鸡口服免疫后可产生CfrA蛋白特异性的血清IgG和肠粘膜sIgA抗体;重组乳酸乳球菌口服免疫后空肠弯曲菌在鸡体内的增殖速度显著低于对照组。【结论】成功构建了重组CfrA蛋白的食品级乳酸乳球菌诱导表达系统;表达CfrA蛋白的重组乳酸乳球菌口服免疫鸡对空肠弯曲菌在鸡肠道的定殖具有一定的抑制作用,为研制重组乳酸菌口服家禽免疫制剂防治空肠弯曲菌奠定了基础。
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| 关 键 词： | 空肠弯曲菌 肠菌素受体CfrA 乳酸乳球菌 表达 口服免疫 |
| 收稿时间： | 2015-11-10 |
| 修稿时间： | 2016-01-14 |
Immunological modulation of chickens by recombinant Lactococcus lactis expressing ferric enterobactin receptor CfrA of Campylobacter jejuni |
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| Chuanwen Wang,Hongzhuan Zhou,Fangfang Guo,Yuxia Chen,Bing Yang,Xia Su,Ruiliang Zhu and Fuzhou Xu. Immunological modulation of chickens by recombinant Lactococcus lactis expressing ferric enterobactin receptor CfrA of Campylobacter jejuni[J]. Acta microbiologica Sinica, 2016, 56(8): 1326-1334 |
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| Authors: | Chuanwen Wang Hongzhuan Zhou Fangfang Guo Yuxia Chen Bing Yang Xia Su Ruiliang Zhu Fuzhou Xu |
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| Affiliation: | Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Technology, Shandong Agricultural University, Tai''an 271018, Shandong Province, China;Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China,Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Technology, Shandong Agricultural University, Tai''an 271018, Shandong Province, China and Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China |
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| Abstract: | |
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| Keywords: | Campylobacter jejuni ferric enterobactin receptor CfrA Lactococcus lactis expression oral immunization |
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