摘要
目的构建表达鼠精子 cyritestin蛋白的可口服减毒沙门氏菌活疫苗株。方法将编码成熟 cyritestin蛋白c DNA序列的酶切片段插入含 p UC复制子的表达载体 p YA3 3 3 9中 ,构建重组质粒 p MCY,将该重组质粒转入鼠沙门氏伤寒杆菌疫苗株 X4 5 5 0 ,获得杂合菌株 X4 5 5 0 ( p MCY)。应用 western blot对重组菌所表达的重组 cyritestin蛋白进行免疫分析。结果该无抗药性的杂合菌株在体外营养选择压力下可较稳定携带重组质粒传代繁殖 ,在体内可较稳定地定居于 Peyer’s结、肠系膜淋巴结和脾脏。该重组菌可高水平表达重组 cyritestin蛋白。结论高效表达 cyritestin蛋白的 X4 5 5 0 ( p MCY)
ObjectiveTo construct a live oral vaccine strain expressing the mature protein of mou se sperm cyritestin using high copy number vector Methods The gene encoding the mature cyritestin protein was obtained by digesting the p lasmid TAZ83/13 using restrictive enzyme and was inserted into the plasmid pYA3339 with a pUC origin The resulting recombinant plasmid pMCY was const ructed and transformed into the delta cya, delta crp, delta asd attenuated S typhimurium vaccine strain X4550, so to get the live vaccine strain X455 0(pMCY) The recombinant cyritestin protein expressed by the X4550(pMCY ) was analyzed by western blot assay Results The recombinant strain X4550(pMCY) is stable and can grow for genera tion s under dietary selection pressure in vitro , and can colonized and persiste d in Peyer's patches, mesenteric lymph nodes for a period The recombinant st rain could express high level of recombinant mature cyritestin protein recognize d by the monoclonal anti cyritestin Conclusions This live vaccine strain X4550(pMCY)are worthy to be studied fo r immunological contraception
出处
《免疫学杂志》
CAS
CSCD
北大核心
2001年第5期344-348,共5页
Immunological Journal
