ISOLATION OF THE ARABIDOPSIS-ABI3 GENE BY POSITIONAL CLONING
ISOLATION OF THE ARABIDOPSIS-ABI3 GENE BY POSITIONAL CLONING
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DOI：
10.1105/tpc.4.10.1251
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发表时间：
1992-10-01
期刊：
影响因子：
11.6
通讯作者：
GOODMAN, HM
中科院分区：
文献类型：
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作者：
GIRAUDAT, J;HAUGE, BM;GOODMAN, HM
拟南芥abi 3突变体在种子发育和萌发的各个方面都发生了改变，这反映了对激素脱落酸的反应性降低。ABI 3基因已通过定位克隆分离。构建了abi 3区域的详细限制性片段长度多态性（RFLP）图谱。一个RFLP标记紧密连锁的abi 3基因座进行了鉴定，并通过分析一组重叠的粘粒克隆含有该标记，abi 3基因座被定位在一个35 kb的区域内。一个11-kb的亚片段，然后显示，以补充转基因植物中的突变表型，从而进一步界定的位置的基因座。在该片段中鉴定出候选ABI 3基因在发育中的果实中表达。从相应cDNA克隆的序列分析中推导出编码蛋白的一级结构。在最严重的abi 3 -4等位基因中，由于存在引入提前终止密码子的点突变，该预测蛋白的大小减少了40%。预测的ABI 3蛋白显示离散区域的玉米胎生-1蛋白的高度相似性。
Arabidopsis abi3 mutants are altered in various aspects of seed development and germination that reflect a decreased responsiveness to the hormone abscisic acid. The ABI3 gene has been isolated by positional cloning. A detailed restriction fragment length polymorphism (RFLP) map of the abi3 region was constructed. An RFLP marker closely linked to the abi3 locus was identified, and by analyzing an overlapping set of cosmid clones containing this marker, the abi3 locus was localized within a 35-kb region. An 11-kb subfragment was then shown to complement the mutant phenotype in transgenic plants, thereby further delimiting the position of the locus. A candidate ABI3 gene was identified within this fragment as being expressed in developing fruits. The primary structure of the encoded protein was deduced from sequence analysis of a corresponding cDNA clone. In the most severe abi3-4 allele, the size of this predicted protein was reduced by 40% due to the presence of a point mutation that introduced a premature stop codon. The predicted ABI3 protein displays discrete regions of high similarity to the maize viviparous-1 protein.