作者：Wen L, Tan B, Guo WW.
作者单位：Huazhong Agr Univ
摘要：Transgene copy number has a great impact on the expression level and stability of exogenous gene in transgenic plants, so transgene copy number analysis is identified as one most important task after obtaining transgenic plants. In this paper, TaqMan real-time PCR was used to estimate the copy number of exogenous MAC12.2 and NPTII genes in transgenic precocious trifoliate orange (Poncirus trifoliata [L.] Raf) in order to overcome the limitations of Southern blot analysis, which is labor-intensive, time-consuming, in considerable needs of DNA, etc. We developed a real-time PCR assay which permitted the determination of the copy number of transgene (MAC12.2 and NPTII), relative to a conserved endogenous gene (PtLTP) in transgenic lines. R value is 0.92 by comparing the results to that of Southern blot analysis, indicating a strong correlation coefficient between TaqMan real-time PCR assay and Southern blot method.
关键词：Transgenic citrus; TaqMan real-time PCR; Copy number
文献注录：Wen L, Tan B, Guo WW.. Estimating transgene copy number in precocious trifoliate orange by TaqMan real-time PCR [J]. Plant Cell Tissue and Organ Culture. 2012, 109(2): 363-371.
报/刊名：《Plant Cell Tissue and Organ Culture》，发表于2012 / 第 109(2) 期。
文献类型： [J]
页码：第 363-371 页 / 共9 页