嗜水气单胞菌外膜蛋白基因ompTS的克隆与序列分析

Cloning and sequence analysis of outer membrane protein gene ompTS of Aeromonas hydrophila

  • 摘要: 根据已发表的外膜蛋白基因ompII的核苷酸序列设计引物,从分离自患红底板病的中华鳖的嗜水气单胞菌中扩增得到了ompTS基因.对ompTS基因进行序列分析,发现其与ompII基因的核苷酸序列有83.5%的同源性.ompTS基因最长的开放阅读框(ORF)为1068nt,编码由355个氨基酸组成,分子量为38.9 kDa的蛋白质OmpTS,其氨基酸序列的前20个氨基酸残基可能组成信号肽.由ompTS基因推导的编码氨基酸序列与其它细菌外膜蛋白的氨基酸序列的比较结果,进一步证实细菌外膜蛋白氨基酸序列的N端存在高度保守区.根据序列分析结果推测,ompTS基因很可能是一个新的基因,编码38.9 kDa的嗜水气单胞菌外膜蛋白OmpTS,该蛋白质在膜中极有可能形成孔道,具备与孔蛋白相似的性质.

     

    Abstract: In this paper , a pair of primers were designed according to the nucleotide sequence of the ompII gene of Aeromonas hydrophila reported. With the specific primers , one target fragment about 1kb was amplified from the A . hydrophila genomic DNA via PCR. The target fragment was inserted into the linearized plasmid vector pRSET A. The nucleotide sequence of the inserted fragment , containing ompTS gene , was determined. The longest open reading frame (ORF) of 1068nt was identified and predicted to encode a 3552aa protein OmpTS with the molecular weight of 38. 9 kDa. Hydrophobicity analysis suggested that except for a potential 202amino2acid signal peptide , the protein was hydrophilic. The nucleotide sequence of ompTS gene showed 83. 5 % homology to the sequence of ompII gene. A Genbank search using the BLASTP program found that , the predicted amino acid sequence of the ompTS gene showed substantial identity with those of the outer membrane proteins reported in Escherichia coli , Klebsiella pneumoniae , Salmonella typhi , Serratia marcescens , Photobacterium profundum and Citrobacter freundii. Based on the amino acid sequence comparison , a high degree of amino acid sequence conservation at N2terminal was revealed. These analysis indicated that the ompTS gene is probably a novel structural gene that codes for OmpTS , which most likely forms pores in the outer membrane.

     

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