摘要
以Pseudomonas syringae的基因组为模板,使用褐藻胶裂解酶引物进行PCR扩增,将目的基因克隆至p MD18-T载体后进行测序.结果显示,克隆基因的大小为1137 bp,预测编码含有378个氨基酸残基的蛋白质.对该蛋白质进一步进行生物信息学分析,结果表明,该蛋白质序列与其他菌株来源的褐藻胶裂解酶具有高的相似性,预测本研究克隆的基因编码褐藻胶裂解酶.该褐藻胶裂解酶的理论分子质量为42.5 ku,理论等电点为8.15.采用同源建模法建立P.syringae褐藻胶裂解酶的三维结构,富含螺旋结构.
The genomic DNA of Pseudomonas syringae was used as the template for amplification of algi- nate lyase gene by PCR using a pair of specific primers. The target gene was cloned into pMD18-T vector and then sequenced. The results showed that the cloned gene was 1137 bp, encoding 378 amino acid residues. This protein was further analyzed by bioinformatics. The results showed that the target protein sequence shared high identities with the alginate lyase sequences from other bacterial strains. So the cloned gene was predicted to be an alginate lyase. The theoretical molecular weight and pI of the alginate lyase were 42. 5 ku and 8. 15, respectively. The three-dimensional structure of P. syringae the alginate lyase was constructed by homology modeling and presented α-helix rich structure.
出处
《集美大学学报（自然科学版）》
CAS
2015年第3期179-185,共7页
Journal of Jimei University：Natural Science
基金
国家自然科学基金资助项目(31401632)
厦门南方海洋研究中心项目(13GZP004NF10)
福建省优良品种同生物与酶工程重点实验室开放基金项目(M20130905)
