口蹄疫病毒株AF/72结构蛋白VP1克隆及其T细胞表位预测

Cloning and T Cell Epitopes Prediction of Structural Protein Gene of a Foot-and-Mouth Disease Virus Strain AF/72

以AF/72RNA为模板,反转录并扩增目的cDNA,然后与pGEM-T easy体连接并转化JM109菌株,提取的重组质粒用凝胶电泳、PCR和EcoRI酶切法鉴定。应用BIMAS生物学软件对AF/72株结构蛋白VP1的H-2Dd、H-2Kd、H-2Ld、A20限制性T细胞表位进行预测,并对不同限制性的T细胞表位进行对比。结果表明,口蹄疫AF/72株结构蛋白VP1长639bp,编码213个氨基酸。不同限制性的T细胞表位虽然不尽相同,但与已鉴定的T细胞表位相比较,它们之间也有一些类似的、并且可能是通用型的T细胞表位。本试验通过生物信息学方法预测AF/72结构蛋白VP1的T细胞表位,为进一步试验确定T细胞表位和口蹄疫病毒T细胞表位的研究提供了一种有效的方法。同时为口蹄疫病毒表位疫苗的设计提供有效地理论基础。

Virus strain AF/72 RNAs were used as templates for RT-PCR. The amplified cDNA products were cloned into pGEM-T easy vectors and transformed into JM109. The recombinant plasmids were identified by electrophoresis,PCR,and EcoR I cleavage. Application of biological software to H-2Dv,H-2Kv,H-2Lv,A20 restricted T cell epitope and the difference of restrictive T-cell epitope were compared. The results showed that the VP1 region of FMDV AF /72 strain was 639 bp in length and codes for a polypeptide of 213 amino acids. Although different restrictive T cell epitope was not identical,but the predicted T cell epitope was similar to compared with identified T cells and might is the universal T-cell. This study prediction of T cell epitope of structural protein of FMDV AF /72 strain through bioinformatics. It would be helpful for identification of T cell epitopes for structural protein of FMDV AF /72 strain using experimental methods and provides an effective method for the virus T cell epitope research simultaneously,for the virus epitope based accines design provides effective theoretical basis.