---

作 者 ：陈灵艳, 陈先进, 蒋烨, 吕暾

期 刊 ：生物技术通报 2016年 32卷 5期 页码：140-145

关键词：猪圆环病毒衣壳蛋白；枯草芽孢杆菌；实时定量PCR；密码子优化；

Keywords:porcine circovirus capsid protein, Bacillus subtilis, RT-qPCR, codon optimization.,

摘 要 ：研究猪圆病毒衣壳蛋白在枯草芽孢杆菌中的表达情况。以3个不同的质粒为基础，构建不同类型的猪圆环病毒衣壳蛋白基因（PCV2-ORF2）重组表达载体，并分别转化进入枯草芽孢杆菌168和WB800中。利用SDS-PAGE和Western blot检测目的蛋白的表达，并通过实时荧光定量PCR检测目的基因的转录水平上的表达情况。结果显示，ORF2基因原序列在枯草芽孢杆菌中难以表达，经过密码子优化后，构建的截短ORF2基因表达载体在枯草芽孢杆菌内表达系统中成功表达目的蛋白。密码子优化促进了目的基因的表达。

Abstract:The objective is to study the expression of porcine circovirus capsid protein in the Bacillus subtilis. Based on 3 different plasmids，different types of recombinant vectors for porcine circovirus capsid protein gene（PCV2-ORF2）were constructed，and they were transformed into strain 168 and WB800 of B. subtilis. The expression of the target protein was detected by SDS-PAGE and Western blot，and the mRNA expression of the target gene in B. subtilis was detected through RT-qPCR. The original ORF2 was difficult to be translated in B. subtilis expression system. After the codon optimization，truncated ORF2 gene was successfully expressed as target protein in intracellular expression system of B. subtilis. Thus，the optimization of the codons promoted the expression of the target gene.

---