Yamada Y, Itano N, Zako M, Yoshida M, Lenas P, Niimi A, Ueda M, Kimata K
Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480-11, Japan.
Biochem J. 1998 Mar 15;330 ( Pt 3)(Pt 3):1223-7. doi: 10.1042/bj3301223.
The structure and organization of mouse hyaluronan synthase 1 gene, HAS1 were determined by direct sequencing of lambda phage clones carrying the entire gene and by application of the long and accurate (LA)-PCR method to amplify regions encompassing the exon-intron boundaries and all of the exons. This gene spans about 11kb of genomic DNA and consists of 5 exons and 4 introns. A similarity in the exon-intron organization was found between the genes of mouse HAS1 and Xenopus laevis DG42 which was recently identified as Xenopus hyaluronan synthase. The transcription initiation site was determined by rapid amplification of the cDNA ends (5'-RACE). Position +1 is located 55 nucleotides upstream of the ATG initiation codon. The promoter region of the HAS1 gene has no typical TATA box, but contains a CCAAT box located 190 nucleotides upstream of the transcription initiation site. Further analysis of 1.4 kb of the 5' flanking region revealed several potential binding motifs for transcription factors. This information about the gene structure may be useful for further studies on the promoter activity.
通过对携带完整基因的λ噬菌体克隆进行直接测序，并应用长片段准确（LA）-PCR方法扩增包含外显子-内含子边界和所有外显子的区域，确定了小鼠透明质酸合酶1基因（HAS1）的结构和组织。该基因跨越约11kb的基因组DNA，由5个外显子和4个内含子组成。在小鼠HAS1基因与非洲爪蟾DG42（最近被鉴定为非洲爪蟾透明质酸合酶）的基因之间发现了外显子-内含子组织的相似性。通过cDNA末端快速扩增（5'-RACE）确定了转录起始位点。+1位点位于ATG起始密码子上游55个核苷酸处。HAS1基因的启动子区域没有典型的TATA盒，但在转录起始位点上游190个核苷酸处含有一个CCAAT盒。对5'侧翼区域1.4kb的进一步分析揭示了几个潜在的转录因子结合基序。这些关于基因结构的信息可能有助于进一步研究启动子活性。
