Cotterill S, Lehman I R, McLachlan P
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, UK.
Nucleic Acids Res. 1992 Aug 25;20(16):4325-30. doi: 10.1093/nar/20.16.4325.
We have isolated both cDNA and genomic clones for the 73 kDa subunit of the DNA polymerase alpha primase of Drosophila melanogaster. Analysis of these clones has identified an open reading frame of 1959 bases coding for a protein of 72.5 kDa. Northern analysis has shown the mRNA for the gene to be approximately 2.5 kb, and comparison of the cDNA and the genomic clones shows that the coding region of the gene lacks introns. The 5' end of the transcript has been mapped by primer extension, and the position of the gene in the genome mapped using in situ analysis. Computer analysis has been carried out on both coding and non coding regions of the gene. The protein sequence shows some homology to the analogous subunit in the S. cerevisiae DNA polymerase alpha, however a search of the data banks failed to reveal other homologies, or provide any clues as to the function of the protein. Analysis of the non-coding regions indicates some potential control regions for the gene. The 73 kDa protein has been overproduced, but a preliminary analysis failed to reveal any enzymatic activities.
我们已经分离出了黑腹果蝇DNA聚合酶α引发酶73 kDa亚基的cDNA克隆和基因组克隆。对这些克隆的分析确定了一个1959个碱基的开放阅读框,编码一个72.5 kDa的蛋白质。Northern分析表明该基因的mRNA约为2.5 kb,cDNA与基因组克隆的比较显示该基因的编码区没有内含子。转录本的5'端已通过引物延伸进行定位,该基因在基因组中的位置已通过原位分析进行定位。已对该基因的编码区和非编码区进行了计算机分析。该蛋白质序列与酿酒酵母DNA聚合酶α中的类似亚基有一些同源性,然而,对数据库的搜索未能揭示其他同源性,也没有提供关于该蛋白质功能的任何线索。对非编码区的分析表明该基因有一些潜在的控制区。73 kDa的蛋白质已过量产生,但初步分析未能揭示任何酶活性。