Physical mapping of unique nucleotide sequences on identified rice chromosomes
Physical mapping of unique nucleotide sequences on identified rice chromosomes
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DOI：
10.1023/a:1006062905742
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发表时间：
1998-12-01
影响因子：
5.1
通讯作者：
Fukui, K
中科院分区：
文献类型：
--
作者：
Ohmido, N;Akiyama, Y;Fukui, K
结合染色体客观鉴定和高灵敏度荧光原位杂交技术（FISH），建立了一种染色体特异区域上独特核苷酸序列的物理定位方法。从水稻基因组DNA中克隆了4个大小在1.3 ~ 400 kb之间的独特核苷酸序列，并将其定位在水稻染色体图谱上。将含有水稻基因组DNA 399 kb插入片段的酵母人工染色体（YAC）克隆定位于水稻染色体长臂（1q2.1）的远端，将含有水稻抗叶瘟基因（Pi-b）的细菌人工染色体（BAC）克隆（180 kb）定位于第2染色体长臂（2q2.1）的远端。将含有抗白叶枯病基因（Xa-21）的粘粒（35 kb）定位于第11号染色体长臂的间隙区（11q1.3）。在水稻第3染色体长臂（4q2.1）的远端区域，成功地定位了一个长度为1.29kb的RFLP标记。对于染色体区域内核苷酸序列的精确定位，图像分析是有效的。通过图像分析，BAC克隆定位于特定区域2q2.1：96.16。将该结果与BAC克隆在遗传图谱上的已知位置进行比较，证实了一致性。明确地证明了使用各种探针的FISH方法在小植物染色体上物理定位核苷酸序列的有效性和可靠性。
A physical mapping method for unique nucleotide sequences on specific chromosomal regions was developed combining objective chromosome identification and highly sensitive fluorescence in situ hybridisation (FISH). Four unique nucleotide sequences cloned from rice genomic DNAs, varying in size from 1.3 to 400 kb, were mapped on a rice chromosome map. A yeast artificial chromosome (YAC) clone with a 399 kb insert of rice genomic DNA was localised at the distal end of the long arm of rice chromosome (1q2.1) and a bacterial artificial chromosome (BAC) clone (180 kb) containing the rice leaf blast-resistant gene (Pi-b) was shown to occur at the distal end of the long arm of chromosome 2 (2q2.1). A cosmid (35 kb) with the resistance gene (Xa-21) against bacterial leaf blight was mapped on the interstitial region of the long arm on chromosome 11 (11q1.3). Furthermore a single RFLP marker, 1.29 kb in size, was mapped successfully to the distal region of the long arm of rice chromosome 3 (4q2.1). For precise localisation of the nucleotide sequences within the chromosome region, image analyses were effective. The BAC clone was localised to the specific region, 2q2.1:96.16, by image analysis. The result was compared with the known location of the BAC clone on the genetic map and the consistency was confirmed. The effectiveness and reliability in physically mapping nucleotide sequences on small plant chromosomes achieved by the FISH method using a variety of probes was unequivocally demonstrated.