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目的:结合水稻胚乳生物反应器的优点,将人乳头瘤病毒(HPV)31、33亚型的L1蛋白编码基因导入水稻,构建HPV31 L1、HPV33 L1植物表达载体,最终实现其在水稻胚乳表达系统中的表达。方法:利用生物信息学方法分析并优化合成HPV31及HPV33 L1蛋白编码基因,将其重组于中间载体pMP3和植物表达载体pCAMBIA1300中,分别构建植物双元表达载体pCAMBIA1300-pMP3-HPV-31 L1和pCAMBIA1300-pMP3-HPV-33 L1,采用遗传转化法经根癌农杆菌介导转化水稻TP309种子的愈伤组织,经共培养、潮霉素筛选和分化再生,获得潮霉素抗性植株。结果和结论:构建了HPV31 L1、HPV33 L1的植物表达载体,经根癌农杆菌介导转化水稻TP309种子的愈伤组织,获得转基因植株。
OBJECTIVE: According to the advantages of rice endosperm bioreactor, the L1 protein coding genes of human papillomavirus (HPV) 31 and 33 subtypes were introduced into rice to construct HPV31 L1 and HPV33 L1 plant expression vectors, and finally their expression in rice endosperm expression system In the expression. Methods: The coding genes of HPV31 and HPV33 L1 were analyzed and optimized by using bioinformatics methods. The recombinant plasmid was recombined in the intermediate vector pMP3 and the plant expression vector pCAMBIA1300 to construct the binary vector pCAMBIA1300-pMP3-HPV-31 L1 and pCAMBIA1300 -pMP3-HPV-33 L1. The calli were transformed by Agrobacterium tumefaciens-mediated transformation into callus of TP309 by using genetic transformation method. The callus of hygromycin resistant plants was obtained through co-culture, hygromycin selection and differentiation regeneration. RESULTS AND CONCLUSION: The plant expression vector of HPV31 L1 and HPV33 L1 was constructed. The callus of rice TP309 seeds was transformed by Agrobacterium tumefaciens and the transgenic plants were obtained.