Kang D, Romans P, Lee J Y
Department of Microbiology, Stockholm University, Sweden.
Gene. 1996 Oct 3;174(2):239-44. doi: 10.1016/0378-1119(96)00088-1.
A genomic DNA sequence encoding a basic lysozyme was isolated from the malaria vector mosquito Anopheles gambiae by screening a library with a probe prepared by PCR of reverse transcribed adult RNA. The sequence consists of an upstream region of about 2 kb, a coding region containing three exons and two introns, and a short 3' untranslated region. The coding region indicates that this mosquito lysozyme consists of a signal peptide of 20 residues followed by an 120 aa mature protein which is very similar to other basic lysozymes. The two small introns, 67 and 76 bp, are located at evolutionarily conserved sites. RT-PCR indicated that this gene is expressed abundantly in sugar-fed adults, and at considerably lower levels when females have fed on blood. Although it remains to be seen whether this gene is induced by bacterial infection, the surrounding sequence contains six sequence motifs very similar to the consensus binding sites for a transcription factor similar to NF-kappa B that are found associated with most insect immune response genes. This lysozyme gene maps to division 27 on the left arm of polytene chromosome 2L. An ORF unrelated to any animal protein in current data bases was found at the 5' end of the clone.
通过用由成年蚊子反转录RNA经PCR制备的探针筛选文库,从疟蚊媒介冈比亚按蚊中分离出编码碱性溶菌酶的基因组DNA序列。该序列由一个约2kb的上游区域、一个包含三个外显子和两个内含子的编码区域以及一个短的3'非翻译区域组成。编码区域表明,这种蚊子溶菌酶由一个20个残基的信号肽和一个120个氨基酸的成熟蛋白组成,该成熟蛋白与其他碱性溶菌酶非常相似。两个小内含子,分别为67bp和76bp,位于进化保守位点。RT-PCR表明,该基因在取食糖水的成年蚊子中大量表达,而在雌蚊吸食血液后表达水平显著降低。虽然该基因是否由细菌感染诱导还有待观察,但周围序列包含六个与类似于NF-κB的转录因子的共有结合位点非常相似的序列基序,这些基序与大多数昆虫免疫反应基因相关。该溶菌酶基因定位于多线染色体2L左臂的27区。在克隆的5'端发现了一个与当前数据库中任何动物蛋白都无关的开放阅读框。