角毛壳菌丝切蛋白基因的克隆和特性分析

在构建的角毛壳菌(Chaetomium cupreum)菌丝体cDNA文库中获得丝切蛋白(Cofilin)的ESTs序列片段,为研究角毛壳菌的生物特性,提高其生物防治效果,以角毛壳菌总RNA反转录产物为模板进行PCR扩增,成功获得了Cofilin基因cDNA序列.生物信息学分析结果表明,该基因全长468bp,编码155个氨基酸,编码蛋白理论分子量17kD,等电点是4.99,α-螺旋和无规则卷曲是其主要的二级结构,为亲水性蛋白质.该蛋白有6个位点发生Ser磷酸化,4个位点发生Tyr磷酸化,有4个保守区,ClustalX分析说明该蛋白与柄孢霉(Podospora anserina)和球毛壳菌(C.g lobosum)亲缘关系较近.

Cloning and Character Analysis of Cofilin from Chaetomium cupreum

Cofilingene was obtained from mycelia cDNA library of Chaetomium cupreum.Full length of Cofilin gene was acquired by PCR amplification with reverse RNA template in order to study the biological character and enhance the biocontrol effect.The bioinformatics results showed that the full-length of Cofilin gene was 468 bp length and encoded 155 amino acids.The protein molecular weight was 17 kD and its theoretical isoelectric point was 4.99.Cofilin was hydrophilic and its main secondary structure elements of cofilin were alpha helix and random coil.Cofilin protein included 4conservative regions and 6Ser and 4Tyr phosphorylation site.ClustalX analysis revealed that the amino acid sequence shared high similarity with P.anserinaand C.g lobosum.