Zhu Xue-Feng, Chen Xue-Wei, Li Xiao-Bing, Qian Qian, Huang Da-Nian, Zhu Li-Huang, Zhai Wen-Xue
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Yi Chuan Xue Bao. 2002 Oct;29(10):880-6.
The transformation mediated by Agrobacterium has been successfully applied to rice in recent years. In the previous research we have transferred the Xa21 gene into five rice varieties of China, using Agrobacterium-mediated trasformation. In this study, T-DNA flanking sequences of Xa21 transgenic rice lines were obtained by using thermal asymmetric interlaced PCR (TAIL-PCR). The flanking sequences which are actual rice DNA were identified and located on molecular linkage map developed from a ZYQ8/JX17 double haploid (DH) population. A total of 22 T-DNA flanking rice sequences were isolated. Nineteen of them displayed RFLPs between the two parents, ZYQ8 and JX17, and were mapped on the rice chromosomes, 3, 4, 7, 9, 10, 11 and 12, respectively. The genetic mapping of T-DNA integration sites in Xa21 transgenic rice will benefit the study of position effect and stable inheritance of the transgene Xa21.
近年来,根癌农杆菌介导的转化已成功应用于水稻。在先前的研究中,我们利用根癌农杆菌介导的转化方法,将Xa21基因导入了中国的五个水稻品种。在本研究中,通过热不对称交错PCR(TAIL-PCR)获得了Xa21转基因水稻株系的T-DNA侧翼序列。对这些实际为水稻DNA的侧翼序列进行了鉴定,并定位到由ZYQ8/JX17双单倍体(DH)群体构建的分子连锁图谱上。共分离出22个T-DNA侧翼水稻序列。其中19个在两个亲本ZYQ8和JX17之间表现出限制性片段长度多态性(RFLP),并分别定位在水稻第3、4、7、9、10、11和12号染色体上。Xa21转基因水稻中T-DNA整合位点的遗传定位将有助于研究转基因Xa21的位置效应和稳定遗传。