摘要
通过分子克隆手段获得了变形假单胞菌(Pseudomonas plecoglossicida CGMCC2039)精氨酸脱亚胺酶(arginine deiminase,简称ADI)基因序列,并将扩增的ADI基因片段插入表达载体pET28a中,构建了ADI的重组表达载体。核苷酸序列分析显示该基因开放阅读框为1254bp,编码417个氨基酸。Blast分析显示它与其他假单胞菌属菌种具有很高的同源性。SDS-PAGE结果显示出分子量大小为48.5kDa的明显的蛋白质特异性条带,并具有酶活。该载体的构建为该基因在大肠杆菌中的表达、纯化和抗肿瘤活性研究奠定了基础。
The arginine deiminase （ADI） gene from Pseudomonas plecoglossicida CGMCC2039 was obtained through molecular cloning method, and was inserted into expression vector pET28a to construct the recombinant ADI expression vector. Nuclcotide sequence analysis showed the gene had an open reading frame of 1254 bp, which encoded 417 amino acids. Blast analysis indicated the deduced amino acid sequence had high identity with that from some other Pseudomonas species. SDS-PAGE exhibited an obvious and specific protein band with a mass weight of 48.5 kDa, and the enzyme activity was confirmed in the later assay. Construction of this vector could be a basis for further research of expression, purification and anti-tumor activity of this gene.
出处
《工业微生物》
CAS
CSCD
北大核心
2008年第4期1-6,共6页
Industrial Microbiology
基金
国家重点基础研究发展规划(No 2003CB716008)
国家高技术研究发展计划资助(No 2007AA02Z226)
长江学者创新团队发展计划(IRT0532)资助
关键词
精氨酸脱亚胺酶
序列分析
表达载体构建
arginine deiminase
sequence analysis
construction of expression vector
