南方菜豆花叶病毒外壳蛋白基因的克隆与原核表达

目的]克隆南方菜豆花叶病毒(SBMV)外壳蛋白(CP)基因,并对其进行原核表达。方法]利用RT-PCR方法克隆SBMV CP基因,将CP基因定向插入表达载体pET28a中,构建其表达载体,并转化大肠杆菌BL-21表达重组蛋白。结果]试验成功克隆到大小为799 bp的SBMV CP基因,测序分析发现与NCBI中目标基因的相似度达99%以上;成功构建了该基因的原核表达载体pET28a-SBMVCP,并确定重组蛋白在37℃、1.0 mmol/L IPTG、4 h条件下表达量最大。结论]该研究为SBMV抗体的制备奠定了基础。

Cloning and Prokaryotic Expression of Coat Protein Gene in Southern Bean Mosaic Virus

Objective ] The paper was to clone the coat protein （CP） gene in Southem bean mosaic virus （SBMV） and to express it in pro- karyote. Method] The SBMV CP gene was cloned by the RT-PCR method. The CP gene was inserted into expression vector pET28a to con- struct its expression vector, and transformed into E. coli BL-21 to express the recombinant protein. Result] The SBMV CP gene was cloned successfully with the size of 799 bp, and the sequence analysis showed the similarity of the gene with the target gene in NCBI was more than 99%. The prokaryotic expression vector pET28a-SBMV CP was constructed successfully, and the recombinant protein had highest expression under the conditions of 37 ℃ , 1.0 mmol/L IPTG, 4 h. Conclusion] The study laid a foundation for the preparation of SBMV antibody.