大豆球蛋白G5A3亚基多肽链抗原表位的预测及分段克隆载体的构建

Prediction of antigen epitope of soybean globulin G5A3 polypeptide chain and construction of segmented cloning vector

为进一步探究大豆球蛋白G5亚基中A3多肽链的抗原表位,现利用DNAstar、SOPMA、Disco Tope 2.0 Sever 3个生物信息学软件对其进行抗原表位的预测。结果显示:3个预测软件均包括的序列为^(90)FEK-LQD^(109)、PDI、PETMQQ、^(181)QQQ-EEE^(200)、PDDERK、PKW。在NCBI网站查询A3多肽链的基因序列和蛋白质序列,共960 bp, 320 aa。综合预测结果将A3多肽链重叠分段,经PCR获取分段目的基因,连接转化、PCR和双酶切结果显示重组载体构建成功,为研究大豆球蛋白G5A3多肽链的分段表达和抗原表位的精准筛选提供参考。

In order to further explore the epitope of the A3 polypeptide chain in the G5 subunit of glycinin, three bioinformatics software, DNAstar, SOPMA and Disco Tope 2.0 Sever were used to predict the epitopes. The results showed that the sequences included in the three prediction software were^(90)FEK-LQD^(109)、PDI,PETMQQ,^(181)QQQ-EEE^(200),PDDERK and PKW. The gene sequence and protein sequence of the A3 polypeptide chain were queried on the NCBI website, which were 960 bp and 320 aa in total. According to the comprehensive prediction results, A3 polypeptide chains were overlapped into segments, and the target genes were obtained by PCR. The results of linkage transformation, PCR and double enzyme digestion showed that the recombinant vector was successfully constructed, providing a reference for studying the segmented expression of soybean globulin G5 A3 polypeptide chains and precise screening of antigen epitopes.