Liaoning Normal University, No.1 Liushu South Street, Gan Jing Zi District, Dalian 116029, China.
Appl Biochem Biotechnol. 2013 Mar;169(5):1523-30. doi: 10.1007/s12010-012-9989-9. Epub 2013 Jan 16.
Exogenous fragment sequence and flanking sequence between exogenous fragment and recombinant chromosome of transgenic wheat B72-8-11b were successfully acquired through PCR amplification with cross-matched primers from exogenous genes. Newly acquired exogenous fragment covered the full-length sequence of transformed genes such as transformed plasmid and corresponding functional genes including marker uidA, promoter ubiquitin, lacZ, 1Dx5, and part of sequence of the wheat genome. A specific PCR detection method for transgenic wheat B72-8-11b strain was established on the basis of primers designed according to flanking sequence. The designed primers revealed specific amplification of 132 bp product of transgenic wheat B72-8-11b strain. This method is characteristics of high specificity, high reproducibility, rapid identification, and excellent accuracy for the identification of transgenic wheat B72-8-11b strain.
通过使用交叉匹配引物从外源基因对转基因小麦 B72-8-11b 的外源片段和重组染色体之间的外源性片段序列和侧翼序列进行了成功的扩增。新获得的外源片段覆盖了转化基因的全长序列，如转化质粒和相应的功能基因，包括标记 uidA、启动子泛素、lacZ、1Dx5 和部分小麦基因组序列。根据侧翼序列设计的引物，建立了针对转基因小麦 B72-8-11b 品系的特异性 PCR 检测方法。设计的引物显示出转基因小麦 B72-8-11b 品系的 132bp 产物的特异性扩增。该方法具有高度特异性、高重复性、快速鉴定和对转基因小麦 B72-8-11b 品系鉴定的准确性的特点。
