3种链霉菌纳米区室蛋白encapsulin在白色链霉菌J1074中的表达

Expression of Three Nanocompartmental Shell Proteins(Encapsulin)from Streptomyces in Streptomyces Albus J1074

目的探寻3种链霉菌纳米区室蛋白(encapsulin)基因是否在链霉菌模式菌中成功表达。方法将3种encapsulin基因重组于白色链霉菌Streptomyces albus J1074基因组中,利用亲和层析、超滤、SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)、纳米液相色谱串联质谱(NanoLC-MS/MS)等技术鉴定其表达情况。结果SDS-PAGE结果显示,与野生型菌株相比,构建的重组菌均能表达相对分子质量(Mr)约30×103的差异蛋白,经NanoLC-MS/MS分析,所表达的差异蛋白与3种encapsulin均具有较高的相似度。结论3种encapsulin基因成功在S.albus J1074中实现异源表达,为链霉菌纳米区室蛋白用于高效合成抗生素奠定基础。

Objective To verify whether three types of Streptomyces nanocompartmental shell protein(encapsulin)genes could be successfully heterologouslly expressed in a type strain of Streptomyces.Methods Three encapsulin genes were recombined into the genome of Streptomyces albus J1074,and their expressions were identified by affinity chromatography,ultrafiltration,SDS-polyacrylamide gel electrophoresis(SDS-PAGE)and nano liquid chromatography tandem mass spectrometry(NanoLC-MS/MS).Results SDS-PAGE results showed that three types of recombinant S.albus J1074 strains were all able to express differential proteins with Mr of 30×103 compared to the wild-type strain.These proteins showed high similarity with three encapsulin proteins ensured by NanoLC-MS/MS technique.Conclusion Three types of encapsulin genes were successfully heterologously expressed in S.albus J1074.These results can lay a foundation for the efficient application of Streptomyces nanocompartmental proteins in the synthesis of antibiotics.