小鼠RPE65基因的序列与结构
Sequence and structure of the mouse gene for RPE65.
作者信息
Boulanger A, Liu S, Yu S, Redmond T M
机构信息
Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892-2740, USA.
出版信息
Mol Vis. 2001 Dec 10;7:283-7.
PURPOSE
To determine the genomic organization of the mouse gene for the retinal pigment epithelium (RPE) specific protein RPE65.
METHODS
A genomic clone containing the entire Rpe65 gene was isolated from a mouse genomic P1 library. Fragments of this clone were subcloned and sequenced by automated fluorescent dideoxy DNA sequencing and analyzed. Direct sequencing of PCR amplification products was used to complete the structure. Primer extension analysis was used to determine the transcription start site.
RESULTS
Southern hybridization of restriction digests of mouse genomic DNA reveals a likely single autosomal gene for Rpe65 with no evidence of pseudogenes. Sequence analysis of the mouse P1 clone for Rpe65 and fragments thereof reveals 14 exons distributed over 27 kbp. The transcription start site is located 57 bp upstream of the initiation codon. The protein encoded by the mouse Rpe65 gene is highly conserved when compared with RPE65s from other species.
CONCLUSIONS
RPE65 is a highly conserved protein and it appears that the genes for the mouse and human RPE65s, at least, are also conserved in overall structure.
目的
确定视网膜色素上皮（RPE）特异性蛋白RPE65的小鼠基因的基因组结构。
方法
从小鼠基因组P1文库中分离出包含整个Rpe65基因的基因组克隆。该克隆的片段进行亚克隆，并通过自动荧光双脱氧DNA测序进行测序和分析。利用PCR扩增产物的直接测序来完善结构。采用引物延伸分析来确定转录起始位点。
结果
小鼠基因组DNA酶切片段的Southern杂交显示，Rpe65可能为单常染色体基因，无假基因证据。对小鼠Rpe65的P1克隆及其片段进行序列分析，发现14个外显子分布在27kbp上。转录起始位点位于起始密码子上游57bp处。与其他物种的RPE65相比，小鼠Rpe65基因编码的蛋白质具有高度保守性。
结论
RPE65是一种高度保守的蛋白质，至少小鼠和人类RPE65的基因在整体结构上也是保守的。
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