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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (04): 83-88    DOI:
    
A Simple and Rapid PCR-based Method for Identifying Transgenic Tobacco Plants Carrying a Single Copy of the Integrated Gene
SONG Feng,SUN Min,LUO Ke-ming
Key Laboratory of Ecoenvironments of Three Gorges Reservoir Region, Ministry of Education, School of Life Sciences, Southwest University, Chongqing 400715, China
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Abstract  

A simple, rapid, and low-cost method to identify transgenic tobacco plants carrying a single copy of integrated DNA by one-step genomic polymerase chain reaction was described. The reaction employs one set of primer pairs that amplify the target gene (NPTⅡ) derived from the integrated T-DNA together with the know endogenous single-copy gene (RNR2) as a reference under the same melting temperature in a single PCR. After PCR amplification of the genomic DNA from transgenic tobacco plants, two expected bands were observed. When the band intensity ratio is one, this means that the transformants are carrying a single copy of the integrated gene. It is further observed that the expected 3∶1 Mendelian ratio was obtained in all single-copy T-DNA transgenic lines.



Key wordsTransgenic tobacco plants      Single-copy integrated gene      Polymerase chain reaction (PCR)     
Received: 16 November 2009      Published: 29 April 2010
Cite this article:

SONG Feng, SUN Min, LUO Ke-Meng. A Simple and Rapid PCR-based Method for Identifying Transgenic Tobacco Plants Carrying a Single Copy of the Integrated Gene. China Biotechnology, 2010, 30(04): 83-88.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I04/83

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