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黄粉虫(Tenebrio molitor L.)幼虫中肠编码肽酶的cDNA转录本的序列分析与分子特征

Sequence analysis and molecular characterization of larval midgut cDNA transcripts encoding peptidases from the yellow mealworm, Tenebrio molitor L.

作者信息

Prabhakar S, Chen M-S, Elpidina E N, Vinokurov K S, Smith C M, Marshall J, Oppert B

机构信息

Department of Entomology, Kansas State University, Manhattan, KS, USA.

出版信息

Insect Mol Biol. 2007 Aug;16(4):455-68. doi: 10.1111/j.1365-2583.2007.00740.x.

Abstract

Peptidase sequences were analysed in randomly picked clones from cDNA libraries of the anterior or posterior midgut or whole larvae of the yellow mealworm, Tenebrio molitor Linnaeus. Of a total of 1528 sequences, 92 encoded potential peptidases, from which 50 full-length cDNA sequences were obtained, including serine and cysteine proteinases and metallopeptidases. Serine proteinase transcripts were predominant in the posterior midgut, whereas transcripts encoding cysteine and metallopeptidases were mainly found in the anterior midgut. Alignments with other proteinases indicated that 40% of the serine proteinase sequences were serine proteinase homologues, and the remaining ones were identified as either trypsin, chymotrypsin or other serine proteinases. Cysteine proteinase sequences included cathepsin B- and L-like proteinases, and metallopeptidase transcripts were similar to carboxypeptidase A. Northern blot analysis of representative sequences demonstrated the differential expression profile of selected transcripts across five developmental stages of Te. molitor. These sequences provide insights into peptidases in coleopteran insects as a basis to study the response of coleopteran larvae to external stimuli and to evaluate regulatory features of the response.

摘要

对从黄粉虫(Tenebrio molitor Linnaeus)前中肠、后中肠或整个幼虫的cDNA文库中随机挑选的克隆进行了肽酶序列分析。在总共1528个序列中,92个编码潜在的肽酶,从中获得了50个全长cDNA序列,包括丝氨酸蛋白酶、半胱氨酸蛋白酶和金属肽酶。丝氨酸蛋白酶转录本在后中肠中占主导地位,而编码半胱氨酸蛋白酶和金属肽酶的转录本主要在前中肠中发现。与其他蛋白酶的比对表明,40%的丝氨酸蛋白酶序列是丝氨酸蛋白酶同源物,其余的被鉴定为胰蛋白酶、胰凝乳蛋白酶或其他丝氨酸蛋白酶。半胱氨酸蛋白酶序列包括组织蛋白酶B样和L样蛋白酶,金属肽酶转录本与羧肽酶A相似。对代表性序列的Northern印迹分析表明,所选转录本在黄粉虫五个发育阶段具有差异表达谱。这些序列为研究鞘翅目昆虫中的肽酶提供了见解,作为研究鞘翅目幼虫对外界刺激的反应以及评估该反应的调节特征的基础。

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