黑曲霉EIM-6 pelA基因的克隆及生物信息学分析

目的:克隆黑曲霉EIM-6果胶裂解酶A基因pelA,用于分析果胶裂解酶A的功能与结构。方法与结果:根据GenBank上黑曲霉pelA保守序列设计引物,采用RT-PCR获得黑曲霉EIM-6pelA基因;序列分析表明,pelA基因具有4个内含子,开放读框为1140 bp,编码379个氨基酸残基,含有由20个氨基酸残基构成的信号肽序列;生物信息学分析表明,果胶裂解酶A为具有一定亲水性的稳定酸性分泌蛋白,具有明显的跨膜结构域,β片层结构是该蛋白的主体结构,空间结构是由反平行的β片层结构为基础包围组成的大环,保守功能区域为Pec_lyase_C结构域。结论:克隆获得黑曲霉EIM-6pelA基因,并进行了生物信息学分析,为蛋白质工程改造果胶裂解酶A奠定了基础。

Cloning and Bioinformatics Analysis of Pectin Lyases A Gene peIA in Aspergillus niger EIM-6

Objective： To clone the pectin lyases（PNL） A gene pelA from Aspergillus niger EIM-6 for the study on molecular structure and function of PNL A. Methods ＆ Results： Based on the gene sequence of peIA from the GenBank database, pelA was cloned by RT-PCR from A.niger EIM-6. Sequence analysis showed that pelA gene has four introns, contains 1140 bp nucleotide acids, encoding 379 amino acids, including a signal peptide of 20 amino acids. Bioinformatics showed that A.niger EIM-6 PNL A was stability of hydrophilic secreted protein, with a clear transmembrane area. {3 sheet was the main components of its secondary structure and the parallel 13 sheet structure composed as large ring surrounded. The structures of PNL included a characteristic functional domain of Pec_lyase_C. Conclusion： The pelA gene was cloned and analyzed by bioinformatics for further research of PNL A.