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根据已发表的嗜水气单胞菌气溶素(Aer)毒素基因的序列,设计1对特异性引物,应用PCR技术,扩增GXL3、GXL5、GXL9共3株罗非鱼嗜水气单胞菌的气溶素成熟蛋白基因,克隆到pMD18-T载体上进行序列测定。测序结果表明,3个菌株Aer毒素成熟蛋白的核苷酸序列为1335bp,编码445个氨基酸,其与分离株No.BAA83088的成熟蛋白基因的核苷酸序列及推导的氨基酸序列的同源性分别为90.9%、91.4%、91.4%和96.2%、96.6%、96.6%,具有很高的同源性。应用分子生物软件分析广西分离株GXL9 Aer成熟蛋白,该蛋白无跨膜区,拥有较多的疏水区和抗原位点,等电点为5.5。
A pair of specific primers was designed based on the published sequence of Aer toxins of Aeromonas hydrophila. PCR was used to amplify three strains of Gilaria tilapia such as GXL3, GXL5 and GXL9 The aerosol mature protein gene was cloned into pMD18-T vector for sequencing. The sequencing results showed that the nucleotide sequence of the three strains of toxin toxin was 1335bp, which encoded 445 amino acids. The nucleotide sequence and the deduced amino acid sequence identity of the mature protein gene of isolate No.BAA83088 were respectively It was 90.9%, 91.4%, 91.4% and 96.2%, 96.6% and 96.6%, respectively, with high homology. Molecular analysis of Guangxi GXL9 Aer mature protein using molecular biology software, the protein transmembrane region, with more hydrophobic regions and antigen sites, isoelectric point of 5.5.