Determination of Exogenous Gene Copy Number for Transgenic Wheat B73-6-1
Abstract：To reveal the copy number of exogenous gene HMW-GS in wheat B73-6-1, a real-time PCR technique for estimating transgene copy number in transgenic wheat B73-6-1 was built,so as to provide the basis for the analysis of integration sites and genetic expression of HMW-GS. In this study,the standard plasmid containing exogenous gene HMW-GS was constructed to detect copy number of HMW-GS in transgenic wheat B73-6-1.The SYBR Green I real-time PCR results showed that the molecule number of the exogenous gene HMW-GS were 8. 62 × 1018,4. 34 × 1018,1. 10 × 1018,and the molecule number of the endogenous reference gene wx012 were 6. 27 × 1017,3. 14 × 1017,7. 84 × 1017,respectively in three repeat experiments. The copy number of HMW-GS in transgenic wheat B73-6-1 were 13. 75,13. 84,14. 01 by caculated. So this assay revealed that transgenic wheat B73-6-1 carried 14 copies of the exogenous gene HMW-GS.
Keywords：transgenic wheatB73-6-1copy numberstandardsexogenous gene
Publication Date：2016-04-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：5( 19-22,30 )