Jin W W, Li X, Li Z Y, Ning S B, Ling D H, Song Y C
Key Lab of MOE for Plant Developmental Biology, Wuhan University, Wuhan 430072.
Shi Yan Sheng Wu Xue Bao. 2001 Sep;34(3):163-8.
Using fluorescence in situ hybridization (FISH) with metaphase preparations, we localized a transferred barnase-psl DNA sequence onto chromosomes in 8 rice transgenic plants. All the tested rice transgenic lines showed hybridization signals on the middle and terminal regions of chromosome arms except for those close to centromeres. In two lines, two different integration sites were identified, and the other lines showed only one integration site. With the aid of Southern analysis and expression detection, we found that the barnase tended to show a higher level expression in the lines whose integration sites near the distal regions of chromosomes, while the expression level became lower in the lines whose integration sites near the centromeres. This result suggested a possible relationship between chromosomal location of transgenes and the expression level. However it showed no obvious relationship between copy numbers and expression levels. In most cases, the results of multi-color FISH showed that barnase-ps1 always integrated at the same position on the chrmosome as the reporter genes(pHctinG).
利用中期染色体标本进行荧光原位杂交(FISH),我们将转移的bar基因-psl DNA序列定位到了8株水稻转基因植株的染色体上。除靠近着丝粒的区域外,所有检测的水稻转基因株系在染色体臂的中部和末端区域均显示出杂交信号。在两个株系中,鉴定出了两个不同的整合位点,其他株系仅显示一个整合位点。借助Southern分析和表达检测,我们发现bar基因在整合位点靠近染色体远端区域的株系中往往表现出较高水平的表达,而在整合位点靠近着丝粒的株系中表达水平较低。这一结果表明转基因的染色体定位与表达水平之间可能存在关联。然而,拷贝数与表达水平之间未显示出明显的关系。在大多数情况下,多色FISH结果表明bar基因-ps1总是与报告基因(pHctinG)整合在染色体的同一位置。