水牛MFSD2A基因CDS序列克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Buffalo MFSD2A Gene CDS Sequence

中文关键词: 水牛  主要促进因子超家族成员2a  基因克隆  生物信息学

英文关键词: buffalo  MFSD2A  gene cloning  bioinformatics

中文摘要:

      本研究运用生物信息学方法对水牛主要促进因子超家族成员2a(major facilitator superfamily domain containing 2a, MFSD2A)基因编码序列进行克隆及分析。以水牛MFSD2A预测基因序列(GenBank登录号：XM_006061915.4)为模板成功克隆了水牛MFSD2A基因完整CDS序列,该序列长1572bp,可编码523个氨基酸残基；分子式为C₂₆₆₉H₄₁₅₀N₆₅₂O₇₂₅S₂₃,分子量57.71 ku,半衰期30 h,消光系数71445,脂肪系数105.56,理论等电点(PI)8.2,不稳定系数为34.78,总平均亲水性为0.462,说明水牛MFSD2A属于碱性稳定疏水蛋白质；二级和三级结构分析结果显示,水牛MFSD2A蛋白主要由α-螺旋、β-折叠、衍生片段和无规则卷曲组成,其中α-螺旋占46.65%,β-折叠占4.59%,衍生片段占16.25 %,无规则卷曲占32.5 %；亚细胞定位结果显示,水牛MFSD2A定位于细胞质膜、内质网、细胞核和线粒体的比例分别为60.9%、30.4%、4.3%和4.4%,说明水牛MFSD2A主要分布于细胞质膜中；跨膜结构和信号肽预测分析表明,该蛋白存在10个跨膜域,不存在信号肽,故此蛋白属于跨膜蛋白,不属于分泌蛋白；磷酸化位点预测结果显示,水牛MFSD2A蛋白共有48个磷酸化位点,其中丝氨酸磷酸化位点22个,苏氨酸磷酸化位点21个,酪氨酸磷酸化位点5个。糖基化位点预测结果显示,水牛MFSD2A蛋白存在1个潜在的N-糖基化位点和 77个O-糖基化位点；同源性分析结果表明,水牛MFSD2A的氨基酸序列与黄牛、绵羊、猪、家犬、骆驼和人的同源相似性分别为98.7%、94.1%、95.8%、92.4%、94.8%和88.5%,说明MFSD2A具有较高的保守性。

英文摘要:

      In this study, bioinformatics method was used to clone and analyze the coding sequence of MFSD2A gene. The complete coding sequence of buffalo MFSD2A gene was successfully cloned from buffalo MFSD2A predictive gene sequence (GenBank accession number: XM_006061915.4). The sequence is 1572bp and can encode 523 amino acids. Amino acid sequence analysis shows that the molecular formula is C2669H4150N652O725S23, molecular weight 57.71ku, half-life is 30 h, extinction coefficient is 71445, aliphatic index is 105.56, PI is 8.2, instability index is 34.78, grand average of hydropathicity is 0.462, which belongs to alkaline stable hydrophobic protein. The results of secondary structure analysis showed that buffalo MFSD2A protein was mainly composed of alpha helix and random coil, in which alpha helix accounted for 46.65%, Beta turn accounted for 4.59%, random coil accounted for 32.5%, extended strand accounted for 16.25%, which was consistent with the prediction results of tertiary structure. The results of subcellular localization showed that the proportion of buffalo MFSD2A located in plasma membrane, endoplasmic reticulum, nucleus and mitochondria was 60.9%, 30.4%, 4.3% and 4.4%, respectively, indicating that buffalo MFSD2A was mainly distributed in the plasma membrane. The prediction analysis of transmembrane structure and signal peptide showed that the protein had 10 transmembrane domains and no signal peptide, so the protein belonged to transmembrane protein and did not belong to secretory protein. The results of phosphorylation site prediction showed that there were 48 phosphorylation sites in buffalo MFSD2A protein, including 22 serine phosphorylation sites, 21 threonine phosphorylation sites and 5 tyrosine phosphorylation sites. Glycosylation site prediction results showed that there was one potential N-glycosylation site and 77 O-glycosylation sites in buffalo MFSD2A protein;The results of homology analysis showed that the homology of buffalo MFSD2A with cattle, sheep, pigs, domestic dogs, camels and humans was 98.7%, 94.1%, 95.8%, 92.4%, 94.8% and 88.5%, respectively, indicating that MFSD2A is highly conservative.