口腔医学研究

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (9): 902-906.DOI: 10.13701/j.cnki.kqyxyj.2016.09.003

• 基础研究论著 • 上一篇    下一篇

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

吴家媛1,2,刘建国1,2*,马欣荣3,洪献忠1,吴志刚1,张剑1,,梁文红1,2   

  1. 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
  • 收稿日期:2016-01-25 出版日期:2016-09-26 发布日期:2016-09-26
  • 通讯作者: 刘建国,E-mail:18798680011@163.com
  • 作者简介:吴家媛(1976~ ),女,江苏泰州人,副主任医师,博士,主要从事口腔牙体牙髓病学临床及基础的研究工作。
  • 基金资助:
    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)

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

WU Jia-yuan1,2, LIU Jian-guo1,2*, MA Xin-rong3, HONG Xiang-zhong1, WU Zhi-gang1, ZHANG Jiang1, LIANG Wen-hong1,2   

  1. 1. Affiliated Stomatology Hospital, Zunyi Medical University, Zunyi 563003, China;
    2. Special Key Laboratory of Oral Diseases Research, Institutions of Higher Learning in Guizhou Province. Key Laboratory of Oral Diseases Research, Zunyi 563000, China;
    3. Chendu Institute of Biology, Chinese Academy of Sciences, Chengdu 610040, China.
  • Received:2016-01-25 Online:2016-09-26 Published:2016-09-26

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235

被引次数

1

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。

关键词: 变异链球菌, 霍乱毒素B亚单位, 转基因黄瓜, 农杆菌

Abstract: 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.

Key words: Streptococcus mutans, B subunit of cholera toxin, Transgenic cucumber, Agrobacterium tumefaciens

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