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- Title
Detection method of exogenous gene in transgenic tobacco by real-time fluorescence quantitative PCR.
- Authors
WANG Sheng; XIE Zhi-xun; XIE Li-ji; XIE Zhi-qin; HUANG Li; LUO Si-si; DENG Xian-wen; HUANG Jiao-ling; LIU Jia-bo
- Abstract
[Objective] The SYBR Green I real-time quantitative PCR method for detecting copy number of exogenous gene in transgenic tobacco was establish to privide reference for detecting copy number of exogenous gene in transgenic plant. [Method] By SYBR Green I real-time fluorescence quantitative PCR technique, the copy number of green-fluorescent protein gene (GFP) as exogenous gene in transgenic tobacco was detected, taking endogenous ribonucleotide reductase gene (RNR2) of tobacco as reference gene. Then two standard curve method of relative quantification was establish to detect copy number of exogenous gene in transgenic tobacco. [Result] The standard curves of GFP gene and RNR2 gene were constructed,which were y=-0.2826x+2.1048 and y=-0.2858x+5.6695, respectively, and the correlation coefficients were 0.9994 and 0.9989. The copy numbers of the five putative transgenic tobaccos were 5, 8, 19, 28 and 45, respectively ,and that of non-transgenic tobacco was 0. [Conclusion] The SYBR Green I real-time quantitative PCR method for detecting copy number of exogenous gene is rapid, convenience and accurate, which is applied to screening transgenic plant, and rapid detection and quantitative analysis of exogenous gene expression.
- Publication
Journal of Southern Agriculture, 2015, Vol 46, Issue 5, p745
- ISSN
2095-1191
- Publication type
Article
- DOI
10.3969/j:issn.2095-1191.2015.5.745