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

第一作者： Xu, K. Y.

作者： Xu, K. Y.;Wang, S. H.;Xi, L.;Wang, Q. J.;Dong, C.;Zhang, J. Y.;Qu, S. C.;Zhang, Z.;Xi, L.;Wang, Q. J.;Dong, C.

作者机构：

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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