噬菌体表面展示技术筛选乙型肝炎病毒前S1蛋白结合蛋白基因

Cloning gene of hepatitis B virus PreS1 binding protein by phage display system

目的 以乙型肝炎病毒 (HBV)前S1蛋白为靶蛋白 ,寻找其相应的结合蛋白。方法 应用T7cDNA文库噬菌体展示技术克隆与前S1蛋白具有结合作用的蛋白序列 ,命名为前S1结合蛋白 (PreS1BP)。将推断的氨基酸序列在蛋白质库中进行搜索 ,自GenBank中获得结合蛋白的cDNA和基因组的全长序列。结果 初步确定PreS1BP即为神经胶质瘤抑制基因区候选基因 2 (GLTSCR2 ) ,cDNA长为14 36核苷酸 ,基因位于第 19号染色体长臂 13.3区 (19q13 3) ,长度 114 4 5碱基对 (bp) ,位于 19号染色体 10 4 0 3483～10 4 14 989,PreS1BP基因含有 13个外显子 ,具有 12个内含子。结论 应用T7cDNA文库噬菌体展示技术和生物信息学方法获得了HBVPreS1BP基因。

Objective T7 cDNA phage display system and bioinformatics methods were employed to find the binding protein to the PreS1 protein of hepatitis B virus (HBV). Methods PreS1 protein was coated in ELISA plate as the target protein, and then T7 cDNA library phage display system was used to scan the binding protein or peptide. A piece of cDNA was found to have the function to bind the PreS1 protein, and the product was named as PreS1 binding protein (PreS1BP). Using BLAST in GenBank, the amino acid sequence of PreS1BP was compared in the protein sequence database. Results The amino acid sequence of PreS1BP was identified as a piece of glioma tumor suppressor candidate region gene 2 (GLTSCR2), and the length of cDNA of PreS1BP was proved to be 1436 nt. The gene was located at chromosome 19q arm (19q13.3) with a length of 11445 base pair between 10403483 and 10414989, containing 13 exons and 12 introns. Conclusion HBV PreS1BP gene could be obtained by T7 cDNA phage display system in combination with bioinformatics methods.