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嗜水气单胞菌J-1株丝氨酸蛋白酶基因克隆与序列分析
引用本文:储卫华.嗜水气单胞菌J-1株丝氨酸蛋白酶基因克隆与序列分析[J].水产学报,2004,28(1):84-88.
作者姓名:储卫华
作者单位:南京农业大学农业部动物疫病诊断与免疫重点实验室,江苏,南京,210095
基金项目:江苏省水产“三项工程”项目
摘    要:根据已发表的气单胞菌胞外蛋白酶基因核苷酸序列,设计和合成了一对引物,以嗜水气单胞菌AhJ—1的基因组DNA为模板,通过PCR技术,扩增到约900bp的丝氨酸蛋白酶基因片段,并克隆到质粒载体pGEM—T中进行测序和分析,结果表明扩增的丝氨酸蛋白酶基因片段与已发表的嗜水气单胞菌丝氨酸蛋白酶Ahe2的同源性有87%,扩增片段编码343个氨基酸,推测的分子量为35700,计算机软件分析表明编码的氨基酸有较高的抗原性,可作为核酸疫苗的侯选基因片段。

关 键 词:嗜水气单胞菌  J-1株  丝氨酸蛋白酶  基因克隆  序列分析  水生动物  病原菌  保护性抗原  基因工程疫苗
文章编号:1000-0615(2004)01-0084-05
收稿时间:2014/3/15 0:00:00
修稿时间:2002年10月16

Cloning and sequence analysis of an extracellular serine-protease gene of Aeromonas hydrophila J-1
CHU Wei-hua.Cloning and sequence analysis of an extracellular serine-protease gene of Aeromonas hydrophila J-1[J].Journal of Fisheries of China,2004,28(1):84-88.
Authors:CHU Wei-hua
Institution:Key Laboratory of Animal Disease Diagnostic and Immunology,Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
Abstract:A pair of primers were designed according to the nucleotide sequences of the protease gene of Aeromonas species reported. With the specific primers, one target fragment about 900bp was amplified from Aeromonas hydrophila strain AhJ-1 genomic DNA via PCR. The PCR products were cloned into pGEM-T vector and sequenced. The sequence was compared with the corresponding regions of the GenBank strains and analyzed by DNAstar software. The nucleotide sequence showed 87 % homology to the sequence of Ahe2 gene. The nucleotide sequence was predicted to encode a 343-aa protein with the molecular weight of 35 700 and with high antigenicity. It suggested that the sequence of serine-protease gene can be used as a candidate fragment for DNA vaccine.
Keywords:Aeromonas hydrophila  serine-protease gene  cloning
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