Maddur A A, Liu X, Zhu Y C, Fellers J P, Oppert B, Park Y, Bai J, Wilde G E, Chen M-S
Department of Entomology, Kansas State University, Manhattan, KS 66506, USA.
Insect Mol Biol. 2006 Aug;15(4):485-96. doi: 10.1111/j.1365-2583.2006.00660.x.
Analysis of transcriptomes from the salivary glands and midgut of Hessian fly larvae Mayetiola destructor (say) identified a set of diverse cDNAs that encode proteins with a relatively high percentage (over 10%) of cysteinyl residues. Structural comparison of these putative proteins with known sequences in GenBank revealed that the positions of the cysteinyl residues in the identified proteins were highly conserved within a family of proteinase inhibitors despite very little overall sequence similarity. Phylogenetic analysis sorted this set of cDNAs into five different groups. To determine if these cDNAs indeed encode proteinase inhibitors, recombinant proteins were generated with two cDNAs from two different groups. Biochemical analysis of the recombinant proteins against commercial and insect gut proteinases demonstrated that the recombinant proteins are strong proteinase inhibitors with different specificities. Northern blot and real-time PCR analysis revealed that the different genes were expressed at different developmental stages and in different tissues. The overall results indicated that M. destructor contains a complex of genes that code for proteinase inhibitors which may regulate proteinase activities in different regulatory pathways. The GenBank accession numbers for the cDNAs in this paper were DQ232690 to DQ232718.
对小麦瘿蚊幼虫Mayetiola destructor (say)唾液腺和中肠转录组的分析鉴定出一组多样的cDNA，这些cDNA编码的蛋白质中半胱氨酸残基的比例相对较高(超过10%)。将这些推定蛋白质与GenBank中的已知序列进行结构比较发现，尽管总体序列相似性很低，但所鉴定蛋白质中半胱氨酸残基的位置在蛋白酶抑制剂家族中高度保守。系统发育分析将这组cDNA分为五个不同的组。为了确定这些cDNA是否真的编码蛋白酶抑制剂，用来自两个不同组的两个cDNA生成了重组蛋白。对重组蛋白针对商业蛋白酶和昆虫肠道蛋白酶的生化分析表明，重组蛋白是具有不同特异性的强蛋白酶抑制剂。Northern印迹和实时PCR分析显示，不同的基因在不同的发育阶段和不同的组织中表达。总体结果表明，小麦瘿蚊含有一组编码蛋白酶抑制剂的基因复合体，这些基因可能在不同的调控途径中调节蛋白酶活性。本文中cDNA的GenBank登录号为DQ232690至DQ232718。
