Spiralin-like protein SLP31 from Spiroplasma eriocheiris as a potential antigen for immunodiagnostics of tremor disease in Chinese mitten crab Eriocheir sinensis
Spiralin-like protein SLP31 from Spiroplasma eriocheiris as a potential antigen for immunodiagnostics of tremor disease in Chinese mitten crab Eriocheir sinensis
复制标题
绒毛螺原体螺旋蛋白样蛋白SLP31作为中华绒螯蟹震颤病免疫诊断的潜在抗原
DOI：
10.1007/s12223-010-0036-3
复制
发表时间：
2010-05-01
影响因子：
2.6
通讯作者：
Wang, Wen
中科院分区：
文献类型：
--
作者：
Meng, Qingguo;Gu, Wei;Wang, Wen
中华绒螯蟹螺原体（Spiroplasma eriocheiris）是导致中华绒螯蟹（Eriocheir sinensis）大量死亡的病原微生物，但对其分子生物学特性的研究还很少。我们在此描述了一个螺旋蛋白样蛋白（SLP31）的鉴定。并在大肠杆菌中表达。核苷酸序列分析表明，该克隆具有一个837 bp的开放阅读框架，编码279个氨基酸的蛋白质。SLP31的理论等电点和摩尔质量分别为7.72和31 kDa。SLP31推导的氨基酸序列与其他物种的螺旋素相似，表明该基因可能是螺旋素家族的一员。螺旋体中的TGA密码子不是作为终止信号，而是作为氨基酸色氨酸的密码。克隆SLP31基因后，将其从TGA突变为TGG，并在E. coli中获得SLP31的全基因表达。Western blotting检测SLP31蛋白的免疫反应性，结果表明SLP31蛋白可作为E. sinensis。
Spiroplasma eriocheiris caused massive mortality of Chinese mitten crab Eriocheir sinensis but little is known about the molecular characteristics of this microorganism. We described here the identification of a spiralin-like protein (SLP31) from S. eriocheiris and expression in Escherichia coli. Analysis of the nucleotide sequence revealed that the clone has an open reading frame of 837 bp encoding a protein of 279 amino acids. Theoretical isoelectric point and molar mass for SLP31 are 7.72 and 31 kDa, respectively. The similarity of SLP31 deduced amino acid sequence shared with the spiralin from other species indicated that the gene may be a member of spiralin family. The TGA codon in Spiroplasma serves not as a stop signal but as a code for the amino acid tryptophan. After cloning the SLP31, the gene was site-mutated from TGA to TGG and transcribed in E. coli to full expression of SLP31. The purified recombinant protein was used to determine the immune reactivity by Western blotting which suggests that SLP31 could be a good antigen for immunodiagnostic of tremor disease in E. sinensis.