Fukui K N, Suzuki G, Lagudah E S, Rahman S, Appels R, Yamamoto M, Mukai Y
Division of Natural Science, Osaka Kyoiku University, Kashiwara, 582-8582 Japan.
Plant Cell Physiol. 2001 Feb;42(2):189-96. doi: 10.1093/pcp/pce026.
Cereal centromeres commonly contain many repetitive sequences that are derived from Ty3/gypsy retrotransposon. FISH analysis using a large DNA insert library of wheat identified a 67-kb clone (R11H) that showed strong hybridization signals on the centromeres. The R11H clone contains Ty3/gypsy retrotransposon-related sequences; both integrase and CCS1 family sequences were identified. Subsequently, we isolated additional 23 large-insert clones which also contained the integrase and CCS1 sequences. Based on the number of the integrase repeats in the clones determined by DNA gel blot analysis, we concluded that the retrotransposon-like sequences are tandemly repeated in wheat centromeres in ca. 55-kb interval on average. This conclusion is consistent with the results of FISH analysis on the extended DNA fibers.
谷物着丝粒通常包含许多源自Ty3/gypsy逆转座子的重复序列。使用小麦的大型DNA插入文库进行的荧光原位杂交(FISH)分析鉴定出一个67 kb的克隆(R11H),该克隆在着丝粒上显示出强烈的杂交信号。R11H克隆包含与Ty3/gypsy逆转座子相关的序列;整合酶和CCS1家族序列均被鉴定出来。随后,我们分离出另外23个大型插入克隆,这些克隆也包含整合酶和CCS1序列。基于通过DNA凝胶印迹分析确定的克隆中整合酶重复序列的数量,我们得出结论,类逆转座子序列在小麦着丝粒中以平均约55 kb的间隔串联重复。这一结论与对延伸DNA纤维的FISH分析结果一致。