转基因水稻中外源基因的荧光原位杂交(FISH)分析  

DETECTION AND ANALYSIS OF ALIEN GENES IN TRANSGENIC RICE BY FLUORESCENCE IN SITU HYBRIDIZATION

作　　者：金危危 李霞[1] 李宗芸[1] 宁顺斌[1] 凌定厚[2] 宋运淳[1] 

机构地区：[1]武汉大学植物发育生物学教育部重点实验室,武汉430072 [2]中国科学院华南植物研究所,广州510650

出　　处：《实验生物学报》2001年第3期163-168,共6页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(39900083);教育部高等学校博士点专项科研基金(207980112)

摘　　要：利用荧光原位杂交(FISH),在供试8个转基因水稻株系的染色体上检出了转入的外源基因bar-nase-ps1片段。其中两株系各检出了两个不同的信号位点,而其他株系均只检出一个信号位点。所有染色体着丝粒区都没有杂交信号。结合Southern杂交及表达分析发现,大多数株系中,整合在染色体端部区的barnase基因表现为高水平表达,而靠近着丝粒区的则表现为低水平表达,表明表达水平与转基因整合位置可能存在一定程度的相关性。表达水平与拷贝数无明显相关。本文还利用多色荧光原位杂交(Multi-color FISH)技术,进行了barnase-ps1片段与报道基因pHctinG的共杂交,多数情况下,barnase-ps1片段与报道基因整合在染色体的同一位置。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 trans-genie 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 trans-genes and the expression level. However it showed no obvious relationship between copy numbers and expression levels. In most cases, the results of multicolor FISH showed that barnase-ps 1 always integrated at the same position on the chrmosome as the reporter genes (pHctinG).

关 键 词：转基因水稻 外源基因 荧光原位杂交 FISH分析 

分 类 号：S511[农业科学—作物学]