Gene cloning and functional analysis of glycosaminoglycan-degrading enzyme chondroitin AC lyase from Flavobacterium columnare G4
Gene cloning and functional analysis of glycosaminoglycan-degrading enzyme chondroitin AC lyase from Flavobacterium columnare G4
复制标题
黄杆菌G4糖胺聚糖降解酶软骨素AC裂解酶基因克隆及功能分析
作者：
H. Xie;P. Nie;M. Chang;Y. Liu;W. Yao
首次从鱼类病原菌柱状芽胞杆菌G4中克隆到软骨素AC裂解酶基因cslA。从第一个转录起始位点开始，cslA将2620个核苷酸延伸到3 '区末端。cslA转录物的开放阅读框包含2286个核苷酸，编码762个氨基酸，在n端有16个残基长的信号肽。在大肠杆菌和重组软骨素AC裂解酶（rChonAC）中成功表达cslA基因，并对其裂解活性进行了分析。与硫酸软骨素共聚的酶谱分析显示了rChonAC的裂解活性，以及从培养的F.柱状芽胞杆菌G4的外周间隙中分离的粗天然ChonAC。在天然天然粗多糖中观察到的低水平裂解活性可能是由于该基因在培养条件下的低水平表达所致。该毒力因子的表达及其作用为进一步研究柱状芽孢杆菌的发病机制提供了研究方向。
The chondroitin AC lyase gene, cslA, was cloned for the first time from the fish bacterial pathogen F. columnare G4. From the first transcription initiation site, the cslA extends 2620 nucleotides to the end of the 3′ region. The open reading frame of cslA transcript has 2286 nucleotides encoding 762 amino acids with a 16 residues long signal peptide at the N-terminus. The gene, cslA was then successfully expressed in Escherichia coli and recombinant chondroitin AC lyase, rChonAC was purified, with its lytic activity analyzed. Zymography analysis copolymerized with chondroitin sulphate revealed the lytic activity of rChonAC and also the crude native ChonAC isolated from periplamic space of cultured F. columnare G4. The low level of lytic activity observed in crude native ChonAC may be due possibly to the low level of expression of this gene in the cultured condition. The expression and the role of this virulence factor is of interest for further research on the pathogenesis of F. columnare.
影响因子：
2.9
作者：
W. Huang;L. Boju;L. Tkalec;H. Su;H. Yang;N. Gunay;R. Linhardt;Y. S. Kim;A. Matte;M. Cygler-M.
通讯作者：
W. Huang;L. Boju;L. Tkalec;H. Su;H. Yang;N. Gunay;R. Linhardt;Y. S. Kim;A. Matte;M. Cygler-M.