固始鸡XCL1基因CDS区克隆与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2021.11.014

摘要/Abstract

摘要： 本研究旨在克隆鸡淋巴细胞趋化因子（lymphotactin，XCL1）基因CDS区并探索其生物学特性。试验以固始鸡胸腺组织总RNA为模板，采用RT-PCR技术对该基因的CDS区进行扩增和克隆，并通过生物信息学软件进行相关生物信息学分析。结果表明，固始鸡XCL1基因CDS区长294 bp，编码97个氨基酸。相似性分析结果发现，鸡XCL1基因CDS区序列与牛、山羊、绵羊、人、小鼠、猪和大鼠的相似性分别为53.7%、53.0%、52.9%、51.8%、51.1%、50.9%和50.4%。进化树结果表明，鸡作为非哺乳动物，与哺乳动物亲缘关系较远。XCL1蛋白分子式为C₄₉₁H₈₂₂N₁₅₀O₁₃₂S₆，理论等电点为10.95，属于碱性蛋白；分子质量为11.13 ku，脂肪系数为102.37，不稳定系数为51.44，属于不稳定蛋白，半衰期为30 h；具有跨膜结构（第5-27位氨基酸）和信号肽区域（第1-18位氨基酸），属于亲水性分泌型蛋白。XCL1蛋白存在8个潜在的磷酸化修饰位点和3个潜在的糖基化修饰位点。XCL1蛋白的二级结构由α-螺旋（35.05%）、β-转角（5.15%）、延伸链（26.80%）和无规则卷曲（32.99%）组成，三级结构预测结果与二级结构一致。亚细胞定位分析表明XCL1蛋白主要在细胞外发挥作用；该蛋白有1个位于第31—88氨基酸残基处的SCY保守性结构域；该蛋白能与OXT、PTAFR、GPR132和XCR1等分子形成互作网络。本试验结果为进一步研究鸡XCL1基因功能提供理论参考。

关键词: 固始鸡; XCL1基因; 克隆; 生物信息学分析

Abstract: The aim of this study was to clone the CDS region of lymphotactin (XCL1) gene in chicken and explore its biological characteristics. Using the total RNA of thymus tissue in Gushi chicken as the template, the CDS of the gene was amplified by RT-PCR and cloned, and the related bioinformatics analysis was carried out by bioinformatics software. The results showed that XCL1 gene CDS in Gushi chicken was 294 bp in length and encoded 97 amino acids. The homology analysis showed that the homology of XCL1 gene CDS sequence in chicken with Bos taurus, Capra hircus, Ovis aries, Homo sapiens, Mus musculus, Sus scrofa and Rattus norvegicus were 53.7%, 53.0%, 52.9%, 51.8%, 51.1%, 50.9% and 50.4%, respectively. The results of phylogenetic tree showed that chicken was a non mammal, and XCL1 gene was far from mammalian. The molecular formula of the XCL1 encoded protein was C₄₉₁H₈₂₂N₁₅₀O₁₃₂S₆, the theoretical isoelectric point was 10.95, which was a basic protein. The molecular mass was 11.13 ku, the fat coefficient was 102.37, and the instability coefficient was 51.44, which was an unstable protein, and the half-life was 30 h. It had a transmembrane structure (amino acids 5-27) and a signal peptide region (amino acids 1-18), and was a hydrophilic secreted protein. There were 8 potential phosphorylation modification sites and 3 potential glycosylation modification sites in XCL1 protein. The secondary structure of XCL1 protein was composed of alpha helix (35.05%), beta turn (5.15%), extended chain (26.80%) and random coil (32.99%). The tertiary structure prediction was consistent with the secondary structure. Subcellular localization analysis showed that XCL1 protein mainly functions outside the cell;the protein had a conserved SCY domain at amino acid residues 31-88. The protein could form an interaction network with OXT, PTAFR, GPR132 and XCR1. The results of this experiment provided a theoretical reference for further study of XCL1 gene function in chicken.

Key words: Gushi chicken; XCL1 gene; cloning; bioinformatics analysis

中图分类号:

S831

田慧慧, 丁梦霞, 郭玉洁, 苏阿茹, 孙桂荣, 田亚东, 蒋瑞瑞, 韩瑞丽, 康相涛, 闫峰宾. 固始鸡XCL1基因CDS区克隆与生物信息学分析[J]. 中国畜牧兽医, 2021, 48(11): 4047-4054.

TIAN Huihui, DING Mengxia, GUO Yujie, SU Aru, SUN Guirong, TIAN Yadong, JIANG Ruirui, HAN Ruili, KANG Xiangtao, YAN Fengbin. Cloning and Bioinformatics Analysis of XCL1 Gene CDS in Gushi Chicken[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4047-4054.

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