| 刘清源，向 华，黄 元，等.携带FMDV…DNA疫苗重组减毒沙门氏菌的构建及其安全性和稳定性研究[J].广东农业科学,2015,42(8):108-112 |
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携带FMDV…DNA疫苗重组减毒沙门氏菌的构建及其安全性和稳定性研究 |
| Construction of attenuated salmonella typhimuriumharboring DNA vaccine of FMDV andits safety and stability |
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| DOI： |
| 中文关键词: FMDV VP1基因 DNA疫苗 减毒沙门氏菌 |
| 英文关键词: FMDV VP1 gene DNA vaccine attenuated salmonella |
| 基金项目: |
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| 摘要点击次数: 1393 |
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| 中文摘要: |
| 为探讨携带 FMDV…DNA 疫苗重组沙门氏菌口服免疫可行性，PCR 扩增O型FMDV主要抗原基因
VP1，并连接到真核表达载体 pIRES，将构建的质粒 pIRES-VP1体外转染BHK-21细胞，间接免疫荧光检测VP1
基因的表达。通过氯化钙转化法将重组质粒 pIRES-VP1转入减毒鼠伤寒沙门氏菌2266，构建2266（pIRES-VP1）
重组菌，并以108
、109
、1010
CFU的剂量口服免疫小鼠，2 周后以相同剂量加强免疫，以研究重组菌在小鼠体内的稳
定性及安全性，并通过将重组菌进行体外传代，研究重组质粒在体外的稳定性。研究结果表明108
、109
CFU剂量
免疫小鼠成活率为100%，1010
CFU剂量口服小鼠出现扎堆现象，且行动迟缓。重组菌体外连传8 代以及免疫小鼠
后从脾脏和肝脏分离鉴定表明，重组菌2266（pIRES-VP1）在体内和体外均具有较好的安全性和稳定性。 |
| 英文摘要: |
| To explore the oral immunization feasibility of live-attenuated salmonella typhimurium for FMDVDNA vaccine，the O type FMDV VP1 gene was amplified by PCR and inserted into expression vector pIRES toconstruct pIRES-VP1. The expression of VP1 gene was proved by indirect immunofluorescence assay in BHK-21cell. The recombinant plasmid was transformed into the attenuated salmonella typhimurium 2266 by calciumchloride process. In order to research the stability and security of recombinant bacteria in mice，immuned miceof recombinant bacteria and strengthen the immune after 2 weeks at the dosage of 108，109，1010 CFU by oraladministration to study the stability of recombinant plasmid in vitro by recombinant bacteria in vitro batches. Theresults showed that 108，109 CFU dose immuned mice，the survival rate was 100% by 1010 CFU dose，clusterphenomenon occurred in mice，and mice became slow.Recombinant bacteria after 8 generation and immunemice in vitro isolated f rom the spleen and liveridentification showed that the recombinant bacteria2266（pIRES - VP1） both in vivo and in vitro hadgood stability. |
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