中国兽医科学2024，Vol.54Issue(4)：504-510,7.DOI:10.16656/j.issn.1673-4696.2024.0066
表达AAL蛋白重组减毒猪霍乱沙门菌的构建及生物学特性评价
Construction and biological evaluation of recombinant attenuated Salmonella choleraesuis expressing AAL protein
摘要
Abstract
In order to construct a recombinant attenuated Salmonella choleraesuis expressing the Aleuria aurantia lectin(AAL),the AAL gene was cloned into the prokaryotic expression vector pTrc-His2A.The recombinant plasmid pTrc-AAL was transformed into the attenuated Salmonella cholerae-suis ΔRposΔslyAΔslrp strain by electroporation to obtain recombinant attenuated SalmonellaΔRposΔslyAΔslrp(pTrc-AAL).The expression of AAL protein in the recombinant strain was identified by Western-blot,and the biological characteristics of the strain were evaluated.The results showed that the recombinant attenuated Salmonella ΔRposΔslyAΔslrp(pTrc-AAL)carrying AAL gene was successfully constructed,and AAL protein could be expressed efficiently;the physiological and biochemical charac-teristics of the recombinant strain ΔRposΔslyAΔslrp(pTrc-AAL)were similar with the ΔRposΔslyAΔslrp(pTrc-AAL)strain;Compared with the wild type(WT)strain,the growth rate of the strain ΔRposΔslyAΔslrp(pTrc-AAL)was slower than WT strain in the logarithmic phase.Similarly,the motility and biochemical characteristics of the strain were also not significantly changed compared with the WT strain.However,the strain ΔRposΔslyAΔslrp(pTrc-AAL)was sensitive to the antibiotics ampicillin and amoxicillin.The LD50 results indicated that the LD50of the recombinant strain ΔRposΔslyAΔsslrp(pTrc-AAL)was at least 8.81 X 105 folds higher than that of the WT strain.In conclusion,this study successfully constructed attenuated Salmonella ΔRposΔslyAΔsllrp(pTrc-AAL)expressing the exogenous protein AAL,which provide a foundation for further exploration of the AAL biological function,and development of mucosal vac-cines targeting Mcells.关键词
橙黄网孢盘菌凝集素(AAL)/猪霍乱沙门菌/生物学特性Key words
Aleuria aurantia lectin(AAL)/Salmonella choleraesuis/biological characterization分类
农业科技引用本文复制引用
蒋华正,黄鑫,赵靖,杨珊珊,杨静,王金泉,刘光亮..表达AAL蛋白重组减毒猪霍乱沙门菌的构建及生物学特性评价[J].中国兽医科学,2024,54(4):504-510,7.基金项目
国家自然科学基金区域联合项目(U22A20522) （U22A20522）
