转基因番茄防龋疫苗中外源目的基因拷贝数的检测

上海口腔医学 ›› 2013, Vol. 22 ›› Issue (6): 613-617.

转基因番茄防龋疫苗中外源目的基因拷贝数的检测

白国辉^1,2,刘建国¹,田源¹,陈筑^1,3,白朋元¹,韩琪¹ ,顾瑜¹,管晓燕¹,王海慧¹

(1.遵义医学院贵州省高等学校口腔疾病研究特色重点实验室，2.医学与生物学研究中心，贵州遵义 563000；3.贵阳市口腔医院，贵州贵阳 550002)

收稿日期:2013-04-05 修回日期:2013-05-10 出版日期:2013-04-12 发布日期:2013-04-12
通讯作者: 刘建国，Tel：0852-8609238，E-mail：13087891001@163.com
作者简介:白国辉(1985-)，男，讲师，硕士研究生，E-mail：baiguohui1228@126.com
基金资助:
国家自然科学基金(30160086，81260164)；贵州省科学技术基金[黔科合J字LKZ[2011]41号]；贵州省科技创新团队资助项目、贵州省重点学科建设资助项目；遵义医学院优秀科研团队培育项目[[2012]12号]

Detection of the exogenous gene copy number of the transgenic tomato anti-caries vaccine

BAI Guo-hui^1,2, LIU Jian-guo¹, TIAN Yuan¹, CHEN Zhu^1,3, BAI Peng-yuan¹, HAN Qi¹, GU Yu¹, GUAN Xiao-yan¹, WANG Hai-hui¹

1. The Special Key Laboratory of Oral Diseases Research, Higher Education Institution of Guizhou Province, 2. Research Center for Medicine & Biology, Zunyi Medical College. Zunyi 563000; 3. The Stomatological Hospital of Guiyang City. Guiyang 550002, Guizhou Province, China

Received:2013-04-05 Revised:2013-05-10 Online:2013-04-12 Published:2013-04-12
Supported by:
Supported by National Natural Science Foundation of China (30160086, 81260164), Science and Technical Fund of Guizhou Province (LKZ[2011]41)， Project of Technology Innovation Team in Guizhou Province, Leading Academic Discipline Construction Project in Guizhou Province and Excellent Scientific Research Team Cultivation Project in Zunyi Medical College ([2012]12).

摘要/Abstract

摘要： 目的：采用SYBR Green实时荧光定量 PCR方法检测转基因番茄防龋疫苗中外源目的基因的拷贝数。方法：用本课题组前期构建的重组质粒pEAC10、pEPC10作为标准品，SYBR Green实时荧光定量PCR法对含外源目的基因pacA-ctxB、pacP-ctxB的转基因番茄植株总基因组样本重复检测，取平均值作为目的基因拷贝数。结果：含pacA-ctxB转基因番茄植株外源目的基因的拷贝数为1.3，含pacP-ctxB转基因番茄植株外源目的基因的拷贝数为3.2。结论：构建的转基因番茄植株属低拷贝转基因植物，具有较好的稳定性。

关键词: 转基因番茄, 实时荧光定量PCR, SYBR Green, 拷贝数, Ct 值

Abstract: PURPOSE: To detect the exogenous gene copy number of the transgenic tomato anti-caries vaccine by using the SYBR Green real-time PCR. METHODS: Recombinant plasmid pEAC10 and pEPC10 were used as standard to detect genome samples of exogenous gene pacA-ctxB and pacP-ctxB by SYBR green fluorescent quantitation, then the average value was calculated as gene copy number. RESULTS: The copy number of the transgenic tomato carrying pacA-ctxB was 1.3 and the pacP-ctxB was 3.2. CONCLUSIONS: The transgenic tomato plants which have high stability are low-copy transgenic plants.

Key words: Transgenic tomato, Real-time fluorescence quantitative PCR, SYBR Green, Threshold cycle, Copy number

中图分类号:

R781

白国辉, 刘建国, 田源, 陈筑, 白朋元, 韩琪 , 顾瑜, 管晓燕, 王海慧. 转基因番茄防龋疫苗中外源目的基因拷贝数的检测[J]. 上海口腔医学, 2013, 22(6): 613-617.

BAI Guo-hui, LIU Jian-guo, TIAN Yuan, CHEN Zhu, BAI Peng-yuan, HAN Qi, GU Yu, GUAN Xiao-yan, WANG Hai-hui. Detection of the exogenous gene copy number of the transgenic tomato anti-caries vaccine[J]. Shanghai Journal of Stomatology, 2013, 22(6): 613-617.

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