水牛MFSD2A基因CDS序列克隆及生物信息学分析

Cloning and Bioinformatics Analysis of CDS Sequence of Buffalo MFSD2A Gene

中文关键词: 水牛  主要促进因子超家族成员2a（MFSD2A）  基因克隆  生物信息学

英文关键词: buffalo  major facilitator superfamily domain containing 2a (MFSD2A)  gene cloning  bioinformatics

中文摘要:

      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，克隆了水牛MFSD2A基因完整序列，全长1572bp，可编码523个氨基酸残基；分子式为C2669H4150N652O725S23，分子质量57.71ku，半衰期30h，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、山羊、猪、骆驼、家犬和人的同源相似性分别为98.7%、98.5%、94.0%、93.5%、91.0%和87.0%，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 the major facilitator superfamily domain containing 2a (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 1572 bp and encodes 523 amino acids. The molecular formula is C2669H4150N652O725S23, molecular weight is 57.71 ku, the half-life is 30 h, extinction coefficient is 71445, aliphatic index is 105. 56, PI is 8. 2, instability index is 34. 78, and grand average of hydropathicity is 0. 462, indicating that buffalo MFSD2A belongs to alkaline stable hydrophobic protein. The analysis results of secondary and tertiary structures 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%, extended strand accounted for 16. 25%, random coil accounted for 32. 5%. The subcellular localization results showed that the proportion of buffalo MFSD2A located in plasma membrane, endoplasmic reticulum, nucleus and mitochondrion 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, goat, pigs, camels domestic dogs and humans was 98. 7%, 98. 5%, 94. 0%, 93. 5%, 91. 0% and 87. 0%, respectively, indicating that MFSD2A was highly conservative.