牛支原体新疆分离株P48基因克隆及分子特征分析

doi:10.16431/j.cnki.1671-7236.2021.02.007

摘要/Abstract

摘要： 本研究旨在分析牛支原体P48基因的序列特性、蛋白的结构和功能。根据GenBank中PG45菌株P48基因序列（GenBank登录号：CP002188.1）设计特异性引物，运用Overlap-PCR扩增获得P48基因全长，其目的片段连接到pMD19-T载体上并进行测序，利用生物信息学方法分析和预测其核苷酸序列及其编码蛋白的理化性质、结构功能（信号肽、跨膜结构、磷酸化位点、糖基化位点、二级结构和三级结构）。结果显示，分离株P48基因序列为1 407 bp，与Mb PG45国际标准株的同源性为100%，聚类于一支；P48基因编码467个氨基酸残基，组成一个无跨膜、稳定的亲水性蛋白；P48蛋白存在信号肽，有44个潜在的磷酸化位点和1个糖基化位点，其二级结构是混合型，其中α-螺旋所占比例最高（41.76%），无规则卷曲次之（37.69%）。功能预测显示，在信号转导、胁迫应答、受体等方面该蛋白存在较高概率。本研究成功克隆了分离株P48基因片段，其编码的蛋白是一个稳定可溶、亲水性蛋白，为分析分离株P48基因的特性和功能提供了一定的理论基础和科学依据。

关键词: 牛支原体; P48基因; 克隆; 生物信息学分析

Abstract: The aim of this study was to analyze the sequence characteristics of P48 gene of Mycoplasma bovis isolated strain in Xinjiang,and the structure and function of its encoded protein.Specific primers were designed based on the sequence of P48 gene of PG45 strain (GenBank accession No.:CP002188.1),the sequence of P48 gene was amplified by Overlap-PCR.P48 gene was inserted into pMD19-T vector for cloning and sequencing.Bioinformatics methods were used to predicted and analyzed its nucleotide sequences and amino acid sequences,including the basic physicochemical properties,signal peptide,transmembrane,phosphorylation site,glycosylation site,secondary structure,tertiary structure.The results showed that the sequence length of Mycoplasma bovis isolated strain P48 gene was 1 407 bp,and its homology with Mb PG45 international standard strain was 100%,and it was clustered in one strain.The P48 protein encoded a protein composed of 467 amino acids,no transmembrane structure,which was a stable soluble hydrophilic protein.The P48 protein had a signal peptide,44 potential phosphorylation sites and 3 glycosylation sites.The secondary structure was mixed,and the alpha helix was the most,accounting for 41.76%,followed by the random coil,accounting for 37.69%.Functional prediction showed that the P48 protein had high probability in signal transduction,stress response and receptor.In this study,the isolated strain P48 gene was cloned successfully,and the protein that it encoded was a stable soluble hydrophilic protein.The results provided a theoretical basis for further exploration of its function of P48 gene of Mycoplasma bovis isolated strain.

Key words: Mycoplasma bovis; P48 gene; cloning; bioinformatics analysis

中图分类号:

S828

凌晨, 郝成武, 张飞, 候凤, 贺笋. 牛支原体新疆分离株P48基因克隆及分子特征分析[J]. 中国畜牧兽医, 2021, 48(2): 457-466.

LING Chen, HAO Chengwu, ZHANG Fei, HOU Feng, HE Sun. Cloning and Bioinformatics Analysis of P48 Gene of Mycoplasma bovis Isolated Strain in Xinjiang[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 457-466.

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