Ishimura-Oka K, Nakamuta M, Chu M J, Sullivan M, Chan L, Oka K
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Genomics. 1995 Oct 10;29(3):751-4. doi: 10.1006/geno.1995.9942.
A complementary DNA for glucokinase (GK) was cloned from mouse liver total RNA by a combination of the polymerase chain reaction (PCR) and mouse liver cDNA library screening. Liver- and beta-cell-specific exons 1 were isolated by PCR using mouse and rat genomic DNAs. These clones were then used to screen a mouse genomic library; three genomic clones were isolated and characterized. The mouse GK gene spans over 20 kb, containing 11 exons including a liver- or beta-cell-specific exon 1, which encodes a tissue-specific 15-aa peptide at the N-terminus of the protein. Both types of GK contain 465 amino acid residues. The predicted amino acid sequence of mouse beta-cell-specific GK showed 98 and 96% identity to the rat and human enzymes, respectively; the corresponding values are 98 and 95%, respectively, for the liver-specific GK. Several transcription factor-binding consensus sequences are identified in the 5' flanking region of the mouse GK gene.
通过聚合酶链反应(PCR)和小鼠肝脏cDNA文库筛选相结合的方法,从小鼠肝脏总RNA中克隆了葡萄糖激酶(GK)的互补DNA。使用小鼠和大鼠基因组DNA通过PCR分离出肝脏和β细胞特异性外显子1。然后用这些克隆筛选小鼠基因组文库;分离并鉴定了三个基因组克隆。小鼠GK基因跨度超过20 kb,包含11个外显子,其中包括肝脏或β细胞特异性外显子1,该外显子在蛋白质的N端编码一个组织特异性的15个氨基酸的肽段。两种类型的GK均包含465个氨基酸残基。预测的小鼠β细胞特异性GK的氨基酸序列与大鼠和人类酶的一致性分别为98%和96%;肝脏特异性GK的相应值分别为98%和95%。在小鼠GK基因的5'侧翼区域鉴定出了几个转录因子结合共有序列。