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羧肽酶是昆虫体内重要的消化酶系之一。利用甜菜夜蛾Spodoptera exigua(Hübner)围食膜多克隆抗体免疫筛选草地螟Loxostege sticticalis L.中肠cDNA表达文库,得到编码羧肽酶A的全长cDNA克隆。该cDNA克隆全长1 380 bp(GenBank登录号EU924506),开放阅读框长1 302 bp,编码434个氨基酸,预测分子量和等电点分别为49.1 kDa和9.56。序列含有胰蛋白酶切割位点和催化特征,具有典型的羧肽酶A特性。将该基因与pET30载体重组后,经IPTG诱导,蛋白在大肠杆菌中获得了表达。
Carboxypeptidase is one of the most important digestive enzymes in insects. A full-length cDNA clone encoding carboxypeptidase A was obtained by immunizing the Loxostege sticticalis L. midgut cDNA expression library with the polyclonal antibody against Peritrichum Spodoptera exigua (Hübner). The cDNA clone was 1 380 bp in length (GenBank accession number EU924506). The open reading frame was 1 302 bp in length and encoded 434 amino acids. The predicted molecular weight and isoelectric point were 49.1 kDa and 9.56, respectively. The sequence contains tryptic cleavage sites and catalytic features with typical carboxypeptidase A properties. After recombination of this gene with pET30 vector, it was induced by IPTG and the protein was expressed in E. coli.