【摘要】 对荧光假单胞菌(Pseudomonas fluorescens ATCC13525)香兰素脱氢酶基因vdh进行了克隆、序列分析以及表达。PCR扩增获得了长度为1 449 bp的核苷酸序列,该序列编码含438个氨基酸,分子量约为50 ku的多肽。序列分析表明该基因与GenBank提供的部分已知vdh基因具有高度的同源性。该基因在大肠杆菌DH5α中能高效表达,而在野生型P.fluorescens ATCC13525中本身并不表达出功能。Vdh基因表达产物香兰素脱氢酶(Vdh)在细胞中主要以可溶性蛋白的形式存在。同时研究表明诱导剂IPTG对vdh基因在大肠杆菌中的表达并不起作用。更多还原

【Abstract】 In this paper the vanillin dehydrogenase gene(vdh) of P.fluroescens ATCC13525 was cloned,sequenced and expressed.The vdh gene was obtained by polymerase chain reaction and was 1 449 bp,enconding a peptide of 438 amino acids with a calculated molecular weight of 50 ku.Nucleotide sequence and amino acid sequence analysis showed the high homology with some of vdh genes in GenBank.The vdh gene was successfully expressed in the recombinant E.coli DH5α and the expression product was mainly soluble protein.But there was no detectable Vdh enzyme activity in wild type P.fluroescens ATCC13525.And it was confirmed that inducer IPTG didn’t effect the expression of the vdh gene in the recombinant E.coli strain.更多还原