摘要： 为了提高枯草芽孢杆菌SX3411菌株羊毛硫细菌素Subtilomycin前体基因subA表达产物的抗原性，采用化学合成的方法合成了SubA的3次重复串联体3×subA基因，构建了该基因的原核表达载体并转入大肠杆菌中。结果表明，3×subA基因在大肠杆菌中获得了融合表达。利用SDS-PAGE纯化大肠杆菌中融合表达的3×SubA，免疫家兔后制备抗体anti-3×SubA，该抗体经检测有较高的滴度。通过Western Blotting杂交，检测了Subtilomycin前体多肽SubA的胞内表达特性。结果表明，枯草芽孢杆菌SX3411在30℃培养的条件下进行不同时间取样检测，枯草芽孢杆菌3411约在培养的第10，12小时胞内有较多的SubA表达。综上所述，通过串联表达Subtilomycin前体基因subA所获得的融合表达产物3×SubA具有较好的抗原性，以此获得的抗体anti-3×SubA完全可以用于枯草芽孢杆菌SX3411中Subtilomycin前体检测和分析。

关键词: 枯草芽孢杆菌SX341, 羊毛硫细菌素, Subtilomycin, SubA蛋白表达, 抗体制备

Abstract: In order to improve the antigenicity of the expression product of Subtilomycin precursor gene of Bacillus subtilis SX3411, a three-times repeated tandem 3×subA gene was synthesized by chemical synthesis method. The prokaryotic expression vector of 3×subA was constructed and transferred into Escherichia coli.The results showed that the 3×subA gene was expressed in E. coli. SDS-PAGE was used to purify the E. coli expressed SubA. After immunizing rabbits, the antibody anti-3×SubA was prepared and had a high titer. The intracellular expression of Subtilomycin precursor polypeptide subA was detected by Western Blotting. The results showed that B. subtilis SX3411 was cultured at 30℃ for different time, and there were more subA expressions in the cells of B. subtilis 3411 at about the 10th and 12th hour. In conclusion, the fusion expression product 3×SubA obtained by tandem expression of the Subtilomycin precursor gene subA has good antigenicity, and the obtained antibody anti-3×SubA can be used for the detection and analysis of Subtilomycin precursors in B subtilis SX3411.

Key words: Bacillus subtilis SX3411, Lantibiotics, Subtilomycin, SubA protein expression, Antibody preparation

中图分类号: 

• TS201.3