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水稻拟南芥FUS6同源基因rFUS6的分离与定位

Isolation and mapping of rFUS6, a rice orthologue of Arabidopsis thaliana FUS6.

作者信息

Huang X, Wang X, Jia H, Feng S, Cao K, Sun C

机构信息

Department of Biochemistry, Fudan University, Shanghai, PR China.

出版信息

DNA Res. 1999 Dec 31;6(6):375-9. doi: 10.1093/dnares/6.6.375.

Abstract

COP9 complex is one of the most important components that act in repressing photomorphogenesis in Arabidopsis thaliana. FUS6 has been identified as one of eight subunits of the COP9 complex in Arabidopsis. Using Arabidopsis Fus6 cDNA as a probe, we screened a rice root cDNA library and a rice genomic library. A 1730-bp cDNA was obtained, which has an open reading frame corresponding to 441-amino-acid. This 441 amino acids putative protein has 67% identity with Arabidopsis COP11/FUS6 (AtFUS6) and 40% identity with human GPS1, an AtFUS6 orthologue. So we designated this novel gene as rFUS6. The 6.2-kb genomic sequence of rFUS6 was also obtained. Sequence comparison showed that the rFUS6 gene had six exons and five introns. Sequence inspection of the 5'-flanking region revealed the presence of some potential light-regulated cis-elements such as a G-box, GT-1 binding sites, and a TGACG motif. Southern hybridization with rice total DNA showed that rFUS6 was perhaps a single copy gene. The rFUS6 locus was mapped by hybridization with a rice BAC library membrane and the results showed that rFUS6 had a locus at 16.3 cM of chromosome 1.

摘要

COP9复合体是拟南芥中抑制光形态建成的最重要组成部分之一。FUS6已被鉴定为拟南芥COP9复合体的八个亚基之一。以拟南芥Fus6 cDNA为探针,我们筛选了水稻根cDNA文库和水稻基因组文库。获得了一个1730 bp的cDNA,其开放阅读框对应于441个氨基酸。这个441个氨基酸的推定蛋白与拟南芥COP11/FUS6(AtFUS6)有67%的同一性,与人类GPS1(AtFUS6的直系同源物)有40%的同一性。因此我们将这个新基因命名为rFUS6。还获得了rFUS6的6.2 kb基因组序列。序列比较表明,rFUS6基因有六个外显子和五个内含子。对5'侧翼区域的序列检查发现存在一些潜在的光调控顺式元件,如一个G盒、GT-1结合位点和一个TGACG基序。用水稻总DNA进行Southern杂交表明,rFUS6可能是一个单拷贝基因。通过与水稻BAC文库膜杂交对rFUS6基因座进行定位,结果表明rFUS6在第1染色体的16.3 cM处有一个基因座。

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