表达鼠精子受精素β的减毒沙门菌疫苗株的构建

Construction of the attenuated salmonella typhimurium strain expressing mouse sperm fertilin β subunit

目的 构建表达鼠精子抗原受精素 β亚单位 (Fβ)的可口服减毒沙门菌活疫苗株。 方法 通过酶切从克隆载体TOPO中获得FβcDNA片段 ,将其插入表达载体PYA314 9中 ,构建重组质粒pFFL ,将该重组质粒转入鼠沙门伤寒杆菌疫苗株X4 6 32 ,获得重组菌株X4 6 32 (pFFL) ,行Westernblot分析及初步毒性试验。 结果 此无抗药性的重组菌株X4 6 32 (pFFL)可稳定表达被Fβ 抗体识别的重组Fβ 蛋白 ,口服该菌株无明显毒性作用。 结论 该疫苗株的构建为研究以Fβ 为靶抗原的精子抗原避孕疫苗打下了基础 ,值得进一步行诱导免疫观察。

Objective To construct a live oral vaccine strain expressing mouse sperm fertilin β subunit. Methods The gene encoding the fertilin β subunit was obtained by digesting a TOPO vector using restriction enzymes and was inserted into the vector plasmid PYA3149.The resultant recombinant plasmid pFFL was constructed and transformed into the delta cya,delta crp,delta asd attenuated S.typhimurium vaccine strain X4632 and the live vaccine strain X4632(pFFL) was got.Its expression was identified by Western blot assay and its toxicity to mice was tested by oral administration. Results The recombinant fertilin β subunit protein expressed stably by X4632(pFFL)could recongnized by the anti-fertilin β subunit antibody.The strain had no biological toxicity to mice. Conclusions This live vaccine strain X4632(pFFL)is worthy to be studied as an immune contraceptive vaccine strain using sperm antigen fertilin β subunit.