Construction and characterization of host-vector balanced lethal system of attenuated salmonella choleraesuisΔcrpΔcyaΔasdC78-1 (pYA3493)
10.3969/j.issn.1000-484X.2015.03.015
- VernacularTitle:减毒猪霍乱沙门菌ΔcrpΔcyaΔasdC78-1（pYA3493）宿主-载体平衡致死系统的构建及其生物学特性研究
- Author:
Ke SHANG
;
Junfeng ZHANG
;
Xiangchao CHENG
;
Chunjie ZHANG
;
Yinju LI
;
Guihua CHEN
;
Yunfei YAN
;
Chongkai ZHAI
;
Zhanqin ZHAO
- Publication Type:Journal Article
- Keywords:
Suicide plasmid;
Host-vector balanced lethal system;
ΔcrpΔcyaΔasdC78-1(pYA3493);
Oral live vaccine vector
- From:
Chinese Journal of Immunology
2015;(3):358-363
- CountryChina
- Language:Chinese
-
Abstract:
Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.
