少孢节丛孢菌丝氨酸蛋白酶家族基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Associated Gene of Serine Protease Family of Arthrobotrys oligospora

本研究在国内首次对少孢节丛孢菌新疆分离株(XJ-A1)丝氨酸蛋白酶家族相关基因P12、P186和P233进行了扩增、克隆及测序,对其编码蛋白信号肽、疏水性与亲水性、高级结构、功能域进行预测和分析。结果显示:3个不同的丝氨酸蛋白酶均含有信号肽和天冬氨酸(D)、组氨酸(H)和丝氨酸(S)3个活性位点,表明它们均属于胞外分泌性Subtilase家族;同时,都含有2个潜在的N末端糖基化位点及2个与底物结合的S1区。系统发生分析发现,P12、P186和P233与其他不同地域、不同种属的丝氨酸蛋白酶基因亲缘性较远。本研究为研发预防动物消化道线虫的生物防控奠定了前期基础。

The serine protease P12,P186 and P233 sequence of A. oligospora Xinjiang isolate（ XJ-A1） was amplified with designed specific primers by PCR,and the signal peptide,hydrophobicity and hydrophilicity,secondary structure,tertiary structure and functional domains of the encoded proteins were predicted and analyzed. The results showed that the different serine proteases of three had a signal peptide. All of them contained three active sites of aspartic acid（ D）,histidine（ H） and serine（ S）,which confirmed that P12,P186 and P233 belonged to the extracellular secreted subtilisin-like serine protease family. Moreover,they contained two potential N-terminal glycosylation sites and two substrate-binding S1 pockets. Phylogenetic analysis revealed that P12,P186 and P233 with other different regions,different species of the serine protease gene had a low degree of sequence identities. The present study could lay a good foundation for further develop the biological agent for prevention and control of animal gastrointestinal nematodes.