林蛙嗜水气单胞菌外膜蛋白基因的克隆、分析及蛋白结构预测

Cloning and sequence anlysis of major outer membrane protein gene of Aeromonas hydrophila and prediction of structure

根据GenBank上发表的嗜水气单胞菌外膜蛋白基因的核苷酸序列设计并合成1对特异性引物,采用PCR方法,扩增出与预期设计的740 bp大小相符的片段,将扩增产物连接到pMD18-T载体上,进行序列测定和分析。结果表明,所克隆的目的基因的核苷酸长为740 bp,共编码246个氨基酸。该基因片段与已发表的嗜水气单胞菌外膜蛋白基因核苷酸序列同源性为98.78%,氨基酸同源性为98.78%。利用生物信息学和分子生物学软件,对嗜水气单胞菌外膜蛋白的结构进行预测,结果表明其为外膜孔蛋白。本试验为嗜水气单胞菌外膜蛋白的表达及基因工程疫苗的研制奠定了基础。

A pair of primers was designed according to Aeromonas hydrophila surface protein gene sequences published in the GenBank. Aeromonas hydrophila surface protein gene strain was amplified by PCR,the fragment of 740 bp in long was inserted into the pMD18 T vecter, then sequenced and analysed. The results showed that the product was 740 bp in size,encoding 246 amino acid residues. The homology of nucleotide sequence with published Aeromonas hydropkila surface protein gene was 98.02 % and 99.40 % at amino acid level. The protein sructure prediction showed that the Aeromonas hydrophila surface protein was porin. This results may be used for the development of vaccine.