变异链球菌表面蛋白PAcP与霍乱毒素B亚单位融合基因在转基因番茄中的表达

发布时间：2018-03-15 12:14

本文选题：变异链球菌　切入点：霍乱毒素B亚单位　出处：《上海口腔医学》2014年03期 　论文类型：期刊论文

【摘要】：目的:在获得含编码变异链球菌表面蛋白PAcP和霍乱毒素B亚单位融合基因的转基因番茄原代种子的基础上,应用分子生物学技术检测外源基因在转基因植株中的表达,测定目的蛋白的含量。方法:PCR筛选含编码变异链球菌表面蛋白PAcP与霍乱毒素B亚单位融合基因的转基因番茄植株;提取番茄果实总蛋白,用BCA试剂盒测定番茄果实总蛋白含量;通过Western印迹检测外源蛋白的表达情况,并用ELISA法对外源目的蛋白含量进行测定。结果:PCR扩增分析可见1.6 kb特异性扩增条带,出现特异性条带的植株占总检测植株的55.6%;转基因番茄总蛋白含量为3.93 mg/mL,Western印迹结果显示,在PVDF膜上,约60 kD处出现特异性条带;ELISA测得表达的目的蛋白占番茄可溶性总蛋白的0.18%。结论:含编码变异链球菌表面蛋白PAcP和霍乱毒素B亚单位融合基因的转基因番茄子代植株能有效表达外源蛋白。
[Abstract]:Objective: on the basis of obtaining transgenic tomato primary seeds containing fusion gene of Streptococcus mutans surface protein PAcP and cholera toxin B subunit, the expression of exogenous genes in transgenic plants was detected by molecular biology technique. Methods the transgenic tomato plants containing fusion gene encoding Streptococcus mutans surface protein (PAcP) and cholera toxin B subunit (CTB) were selected and the total protein of tomato fruit was extracted and the total protein content of tomato fruit was determined by BCA kit. The expression of exogenous protein was detected by Western blotting, and the content of exogenous protein was determined by ELISA. The plants with specific bands accounted for 55.6% of the total tested plants, and the total protein content of transgenic tomato was 3.93 mg / mL Western blot. The target protein expressed by Elisa was 0.18% of the total soluble protein in tomato. Conclusion: transgenic tomato progenies containing fusion gene encoding Streptococcus mutans surface protein PAcP and cholera toxin B subunit are obtained by Elisa. The strain can express foreign protein effectively.
【作者单位】：遵义医学院口腔学院;贵州省高等学校口腔疾病研究特色重点实验室;贵阳市口腔医院;
【基金】：国家自然科学基金(30160086) 贵州省科技支撑计划项目[黔科合SY字(2012)3086号] 贵州省第六批科技创新人才团队资助项目[黔科合人才团队(2013)4026号] 贵州省科学技术基金项目[黔科合J字LKZ(2011)41号]~~
【分类号】：R780.2

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