摘要
为了制备酵母菌液泡电导1(Yvc1)的抗原蛋白,本文首先通过生物信息学方法预测Yvc1抗原表位,并克隆其抗原编码基因。利用NCBI GenBank数据库、ORP Finder、EXPASY ProtScale、SOPMA、IEDB和NetMHCⅡpan等生物信息学工具对酿酒酵母S288c的Yvc1抗原表位进行预测,并根据预测的结果设计引物并克隆其抗原编码基因。结果显示,YVC1基因全长2028 bp,有33个开放阅读框,最长一个被通读的序列,编码675个氨基酸。Yvc1分子量7.7937×10^(4),属于稳定、亲水性蛋白质;该蛋白含有4个膜外区域,亚细胞定位于液泡膜;二级结构中的无规则卷曲和β转角分别占29.48%和3.11%;分别含有5个B细胞优势抗原表位和2个Th优势表位区域;最终预测,优势抗原表位在1~236位氨基酸区域。扩增的酿酒酵母DL5168抗原编码基因序列与预测抗原的基因序列487707~489734 bp位点的相似性达到99%。本研究表明,生物信息学方法成功预测并克隆到Yvc1抗原编码基因,这为其抗原蛋白制备奠定基础。
For the preparation of the antigen protein of yeast vacuolar conductance 1(Yvc1),the bioinformatics methods were applied in the prediction for antigen epitope of Yvc1,and the corresponding antigen encoding gene was cloned in this study.The prediction for antigen epitopes of Yvc1 in Saccharomyces cerevisiae S288c was conducted by using NCBI GenBank database and various bioinformatics software,such as ORP Finder,EXPASY-ProtScale,SOPMA,IEDB,NetMHCⅡpan,etc.The results showed that YVC1 gene contained 2028 bp in length and 33 open reading frames,with the longest one encoding 675 amino acids.As a stable and hydrophilic protein with molecular weight of 7.7937×10^(4),Yvc1 was located in vacuole membrane with 4 extramembrane regions.Regarding the secondary structure,29.48%and 3.11%amino acids were involved in the formation of random coil andβ-turn,respectively.Moreover,our prediction showed there were 5 potential dominant linear B cell epitopes and 2 potential dominant helper T cell epitopes in this protein,with the dominant antigen epitope in the amino acid sequence from 1 to 236.Notably,the gene sequence similarity reached 99%between the amplified antigen encoding gene in YVC 1 of S.cerevisiae DL5168 and the predicted gene from 487707-489734 bp in the strain S288c.Overall,the antigen encoding gene was successfully cloned according to the predicted results by bioinformatics methods,providing a foundation for further preparation of Yvc1 antigen protein in S.cerevisiae.
作者
董晓宇
邱宇
曹钧雄
DONG Xiaoyu;QIU Yu;CAO Junxiong(College of Life and Health,Dalian University,Dalian 116622,China)
出处
《生物加工过程》
CAS
2024年第3期278-288,共11页
Chinese Journal of Bioprocess Engineering
基金
国家自然科学基金(21476032)。
