农业生物技术学报2011，Vol.19Issue(2)：221-229,9.DOI:10.3969/j.issn.1674-7968.2011.02.004
不同转化方法获得的转基因棉花外源基因拷贝数分析
Comparative Analysis of Exogenous Gene Copy Numbers in Transgenic Cotton Transformed by Different Methods
摘要
Abstract
Identifying the copy number of foreign gene in transgenic primary plants is the basic step in genetic engineering application. In this paper, Real-time PCR and the related PfaffI method, usually applied to studying transgenie expression,was used to analyze comparatively the differences of exogenous gene copy number of transgenic cotton (Gossypium hirsutum) T0 plants transformed with one same vector by three different methods,Agrobacterium-mediated, gene-gun bombardment and pollen tube injection, respectively. Results showed that in Agrobacterium-mediated T0 generation, the transgenic plants integrated one or two copy number accounted for only 26.2％ of the total plants, respectively; In contrast, that of transgenic plants transformed by gene-gun bombardment and pollen tube injection were 60％ and 42.8％, respectively. Transgenic lines with three to five copies accounted for 47.5％, 40％ and 57.1％, respectively. In addition, in Agrobacterium-mediated T0 generation, transgenic lines with more than 5 copies accounted for 27.3％, while there was no transgenic plant with more than 5 copies in genetically modified T0 generation transformed by gene gun bombardment or pollen tube methods. These results provide some rationale for selection of transformation methods in crops transgene breeding.关键词
遗传转化方法/转基因棉花/实时定量PCR/转基因拷贝数差异Key words
Genetic transformation methods/ Transgenic cotton(Gossypium hirsutum L)/ Real-time quantitabive PCR/ Difference of exogenous gene copy number引用本文复制引用
杨晓杰,刘传亮,张朝军,武芝霞,张雪妍,刘坤,房卫平,李付广..不同转化方法获得的转基因棉花外源基因拷贝数分析[J].农业生物技术学报,2011,19(2):221-229,9.基金项目
本研究由国家重点基础研究发展规划(973)项目(2010CB126005)资助 （973）
