PRRSV-GP5基因的克隆及其B细胞抗原表位预测与分析

摘要

【目的】研究PRRSV-GP5基因B细胞抗原表位性质。【方法】根据GenBank上发表猪繁殖与呼吸综合征病毒（PRRSV）ATCC VR-2332的全基因序列，设计并合成引物，采用RT-PCR技术，对阳性病料扩增回收，并将其克隆入pMD18-T载体中，送上海生工测序。应用生物信息学同源模型化的方法建立其PRRSV-GP5蛋白3D结构，并根据生物信息学软件DNAStar预测PRRSV-GP5蛋白的二级结构、表面可及性、抗原指数、亲水性及柔韧性，综合分析预测其B细胞抗原表位。【结果】结果表明，其肽段30-36、50-55、140-142、146-151和196-198可能是其优势B细胞抗原表位区域。【结论】该基因高度保守，PRRSV-GP5蛋白呈现较规则的空间结构，同时该结果将为PRRSV-GP5蛋白体外表达产物的应用和基因工程疫苗的研究提供理论依据。%Objective]The objective of this experiment was to study the B cell epitopes of PRRSV-GP5 gene.[Method]Primers for RT-PCR amplification of PRRSV isolates from Heilongjiang were designed according to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published in the GenBank,and sequences were cloned into pMD18-T vector respectively and sequenced in Shanghai. The 3D model of PRRSV-GP5 protein was obtained by homology modeling of bioinfor-matics,and DNAStar was used to predict the secondary structure,surface probability,antigenic index, hydrophilicity and flexibility region.[Result]The peptides of PRRSV-GP5 including 30-36,50-55, 140-142,146-151 and 196-198 were larvaceous dominant peptide.[Conclusion]The results indicat-ed that the target gene of PRRSV-GP5 was highly conservative and space structure of PRRSV-GP5 was regular. The results will provide theoretical evidence for application of expression product and engineering vaccine of PRRSV-GP5 protein.