Voll W, Voit R
Zoologisches Institut, Universität München, Federal Republic of Germany.
Proc Natl Acad Sci U S A. 1990 Jul;87(14):5312-6. doi: 10.1073/pnas.87.14.5312.
The gene for the hemocyanin subunit e of the tarantula Eurypelma californicum has been isolated from a genomic phage library by using a corresponding cDNA clone as a probe. The transcriptional unit spans a chromosomal region of about 55 kilobase pairs (kbp). The gene consists of nine exons that are separated by large introns. The intron/exon boundaries were determined by direct comparison of genomic and cDNA sequences. A putative promoter region ("TATA" box, reversed "CAAT" box) 100 bp 5' to the translational initiation codon strongly suggests the presence of a functional gene. The 3' flanking region carries the polyadenylylation signal (AATAAA) and several conserved structures for the 3' splicing of the pre-mRNA. A comparison of the gene architecture of the subunit e gene with the three-dimensional structure of the arthropod hemocyanin subunit shows a good correspondence with the division of the subunit into three domains (two exons coding for the first, three coding for the second, and four coding for the third domain). The relationship to molluscan hemocyanins, different tyrosinases, and the larval serum proteins is discussed.
通过使用相应的cDNA克隆作为探针,从狼蛛(Eurypelma californicum)的基因组噬菌体文库中分离出了血蓝蛋白亚基e的基因。转录单元跨越约55千碱基对(kbp)的染色体区域。该基因由九个外显子组成,这些外显子被大的内含子隔开。内含子/外显子边界通过对基因组和cDNA序列的直接比较来确定。在翻译起始密码子上游100 bp处的一个假定启动子区域(“TATA”框、反向“CAAT”框)强烈表明存在一个功能基因。3'侧翼区域携带多聚腺苷酸化信号(AATAAA)和几个用于前体mRNA 3'剪接的保守结构。将亚基e基因的基因结构与节肢动物血蓝蛋白亚基的三维结构进行比较,结果表明该亚基分为三个结构域(两个外显子编码第一个结构域,三个编码第二个结构域,四个编码第三个结构域),二者具有良好的对应关系。文中还讨论了与软体动物血蓝蛋白、不同酪氨酸酶和幼虫血清蛋白的关系。