Abstract:In order to clone the gene of aminopeptidase N (APN), the receptor of Bt insecticidal crystal protein of Culex pipiens pallens, and analyze the biological information of the coding protein, single mosquito larva RNA was extracted from Cx. pipiens pallens and the APN cDNA sequence was amplified by RT-PCR. The PCR products were cloned into the pMD-19T simple vectors and sequenced. The sequence homology and the structural feature of the coding protein were analyzed by biological information systems. Results showed that the full-length of CpAPN was 1 383 bp that encoded a protein of 460 amino acid residues. The homology with Culex quinquefasciatus (XM_001862270.1) was 100%. The predicted molecular weight and isoelectric point were 52.9 kDa and 6.33, respectively. It contained an N-terminal signal peptide with 21 amino acids, and two N- glycosylaion sites (93NLT95and 169NVS171), showing the typical characteristics of peptidase family M1, and with the zinc-binding motif HEXXH and the conserved GAMAN motif. Aminopeptidase N gene of Culex pipiens pallens was cloned successfully and analyzed by bioinfomatics technique. The results provided experimental evidence for revealing the function of CpAPN and the insecticidal mechanism of Bt toxin.
郭秀霞, 程鹏, 杨培培, 王海防, 刘丽娟, 张崇星, 王怀位, 公茂庆. 淡色库蚊氨肽酶N基因的克隆及生物信息学分析[J]. 中国人兽共患病学报, 2016, 32(3): 240-245.
GUO Xiu-xia, CHENG Peng, WANG Hai-fang, LIU Li-juan, YANG Pei-pei, ZHANG Chong-xing, WANG Huai-wei, GONG Mao-qing. Molecular cloning and bioinformatics analysis on aminopeptidase N of Culex pipiens pallens. Chinese Journal of Zoonoses, 2016, 32(3): 240-245.
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