摘要
为证明原位整合能否用于枯草芽孢杆菌芽孢表面展示外源蛋白,本研究将芽孢衣壳蛋白CotB、CotZ的编码序列(不含终止密码子),与绿色荧光蛋白基因重组,构建含有融合片段cotB-gfp和cotZ-gfp的原位整合型表面展示载体pEYW25、pEYW26;将上述质粒分别转入枯草芽孢杆菌(Bacillus subtilis PY79)获得重组菌株BSYW25和BSYW26。荧光显微镜下观察上述重组菌的芽孢,只有BSYW26能检测到荧光,Western blot结果显示BSYW26的芽孢外壳蛋白中含有GFP。结果表明原位整合作为一种新的表面展示方法能够应用于枯草芽孢杆菌芽孢表面展示。
To prove whether single crossover integration can be used in the spore surface,the single crossover integration vector pEYW25 harboring fusion fragment cotB-gfp and pEYW26 harboring fusion fragment cotZ-gfp were constructed by fusing the green fluorescence gene to the coding gene(not including the termination codon)of spore coat proteins CotB and CotZ.Recombinant strains BSYW25 and BSYW26 were obtained by transforming pEYW25 and pEYW26 to Bacillus subtilis PY79.The spores of the recombinant bacteria were observed under fluorescence microscope,only the green fluorescence of recombinant strain BSYW26 was detected.Western blot results show the spore coat protein of BSYW26 contains GFP.The results showed that single crossover integration as a new surface display method can be used in Bacillus subtilis spore surface display.
作者
杨亚威
杨明明
龚月生
YANG Yawei;YANG Mingming;GONG Yuesheng(College of Animal Science and Technology,Northwest A&F University,Yangling Shaanxi,712100)
出处
《家畜生态学报》
北大核心
2018年第10期10-15,共6页
Journal of Domestic Animal Ecology
基金
国家自然科学基金(31372343)
关键词
芽孢
原位整合
表面展示
spore
in-situ integration
surface display
