变异链球菌表面蛋白A区与霍乱毒素B亚单位嵌合基因转基因黄瓜植株的获得及鉴定  

Obtain the Transgenic Cucumber of the Fusion Gene of Region A of PAc Gene of Streptococcus mutans and B Subunit of Cholera Toxin

作　　者：吴家媛[1,2] 刘建国[1,2] 马欣荣 洪献忠[1] 吴志刚[1] 张剑[1] 梁文红[1,2] 

机构地区：[1]遵义医学院附属口腔医院牙体牙髓病科,贵州遵义563099 [2]贵州省高等学校口腔疾病研究特色重点实验室,遵义市口腔疾病研究重点实验室,贵州遵义563099 [3]中科院成都生物所,四川成都610040

出　　处：《口腔医学研究》2016年第9期902-906,共5页Journal of Oral Science Research

基　　金：国家自然科学基金资助项目(编号:30160086,39770797);贵州省科技创新人才团队资助项目(合同号:黔科合人才团队[2013]4026);贵州省高等学校特色重点实验室建设项目(合同号:黔教合KY字[2013]109);省市科技合作专项资金(合同号:省市科合【2014】41号)

摘　　要：目的:经农杆菌介导将变异链球菌表面蛋白A区与霍乱毒素B亚单位嵌合基因导入黄瓜,获得转基因黄瓜植株,为转基因可食防龋疫苗的研究提供实验基础。方法:利用双元载体pCAMBIA2301构建变异链球菌表面蛋白A区与霍乱毒素B亚单位嵌合质粒(PAcA-ctxB)的植物表达载体p2355-PAcA-CTB;电转化法导入农杆菌EHA105;采用叶盘转化法转化黄瓜,转基因植物经过卡那霉素抗性筛选、GUS基因染色、PCR及Southern blot杂交分析检测目的基因鉴定。结果:成功得到转基因黄瓜植株,卡那霉素抗性筛选、GUS基因染色、PCR及Southern blot杂交分析检测证实目的基因PAcA-ctxB已整合至黄瓜基因组中。结论:获得了含有变异链球菌表面蛋白A区与霍乱毒素B亚单位嵌合基因的转基因黄瓜植株,为转基因可食防龋疫苗的进一步研究提供了实验基础。Objective：To establish cucumber regeneration system,for the fusion gene of region A of pac gene of S.muant and B subunit of cholera toxin so as to provide useful information for further study of edible vaccine against caries.Methods：The plasmid pCAMBIA2301 was used to construct the recombinant plant expression plasmids p2355-PAcA-CTB.The recombinant plasmids were transformed into Agrobacterium tumefaciens EHA105 by electroporation.Cucumber was transformed by co-cultivating leaf discs with Agrobacterium strains harboring p2355-PAcA-CTB.The regenerated kanamycin-resistant plants were analyzed by PCR and Southern blot.Results：The regenerated transgenic cucumber system was obtained successfully.The results of PCR and Southern blot indicated that the fusion gene of streptococcus mutans A region and the CTB B submit（PAcA-ctxB）was integrated into the genomic DNA of the cucumber.Conclusion：Transgenic cucumbers carrying the fusion gene of streptococcus mutans A region and the CTB B submit were obtained.

关 键 词：变异链球菌 霍乱毒素 B亚单位 转基因黄瓜 农杆菌 

分 类 号：R781.1[医药卫生—口腔医学]