Rapid determination of transgene copy number in tobacco by competitive PCR using a pair of SSR primers
文献类型： 外文期刊
作者： Xu, K. Y. 1 ; Wang, S. H. 1 ; Xi, L. 1 ; Wang, Q. J. 1 ; Dong, C. 1 ; Zhang, J. Y. 1 ; Qu, S. C. 1 ; Zhang, Z. 1 ;
作者机构： 1.Nanjing Agr Univ, Coll Hort, Nanjing, Jiangsu, Peoples R China
2.Univ British Columbia, Dept Wood Sci, Vancouver, BC V5Z 1M9, Canada
3.Liaoning Agr Vocat Tech Coll, Yingkou, Liaoning, Peoples R China
4.Heilongjiang Acad Agr Sci, Dept Hort, Haerbin, Heilongjiang, Peoples R China
关键词： Copy number;Transgene;Competitive PCR;SSR;Tobacco
期刊名称：GENETICS AND MOLECULAR RESEARCH （ 影响因子：0.764； 五年影响因子：0.912 ）
ISSN： 1676-5680
年卷期： 2010 年 9 卷 2 期
页码：
收录情况： SCI
摘要： We developed a straightforward, rapid, and inexpensive method to determine transgene copy number in tobacco. The plasmid (pSSRCopy) used for tobacco transformation contains a simple sequence repeat (SSR) locus, PT1199, which was partially deleted in the middle, a homogenous SSR locus in tobacco K326. A 168-bp segment of the cloned PT1199 was shortened to 95 bp by deleting a 73-bp internal fragment. Using a pair of SSR primers, competitive PCR was amplified from genomic DNA from transgenic tobacco harboring pSSRCopy, and the two expected bands were found. The 168-bp band (SSR-168) corresponds to endogenous PT1199 and the 95-bp band (SSR-95) comes from the integrated pSSRCopy. A single copy of a transgene can be easily distinguished from multiple copies by comparing band densities.
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