Barillas-Mury C, Wells M A
Department of Biochemistry, University of Arizona, Tucson 85721, USA.
Insect Mol Biol. 1993;2(1):7-12. doi: 10.1111/j.1365-2583.1993.tb00119.x.
A 4.1 kb genomic clone of the late trypsin gene from the mosquito Aedes aegypti was isolated, mapped and subcloned. A 1.6 kb subclone, corresponding to 1.1 kb of upstream regulatory region and 0.5 kb of coding region, was sequenced. The gene has no introns within the coding region. The 5' end of the mature mRNA was mapped using primer extension analysis. A TATA box consensus sequence (TATAAA) was found at position -31 from the 5' end of the mature mRNA. A cluster of five repeat sequences homologous to the yeast GCN4 DNA binding site was found within 200 nucleotides upstream of the cap site. GCN4 is required for derepression mediated control of general amino acid biosynthesis in response to amino acid starvation in yeast. It activates the transcription of at least twenty different genes coding for enzymes involved in amino acid biosynthesis. The presence of this cluster of consensus sequences suggests that a protein similar to GCN4 might regulate expression of the late trypsin gene in the mosquito. Southern blot analysis of genomic DNA indicates that late trypsin is a single copy gene.
从埃及伊蚊中分离、定位并亚克隆了晚期胰蛋白酶基因的一个4.1 kb基因组克隆。对一个1.6 kb的亚克隆进行了测序，该亚克隆对应1.1 kb的上游调控区和0.5 kb的编码区。该基因在编码区内没有内含子。使用引物延伸分析确定了成熟mRNA的5'端位置。在成熟mRNA 5'端-31位发现了一个TATA盒共有序列（TATAAA）。在帽位点上游200个核苷酸内发现了一组与酵母GCN4 DNA结合位点同源的五个重复序列。GCN4是酵母中响应氨基酸饥饿介导的一般氨基酸生物合成去阻遏调控所必需的。它激活至少二十种不同的编码参与氨基酸生物合成的酶的基因的转录。这组共有序列的存在表明，一种类似于GCN4的蛋白质可能调节埃及伊蚊中晚期胰蛋白酶基因的表达。基因组DNA的Southern印迹分析表明晚期胰蛋白酶是一个单拷贝基因。
