Cloning, expression, purification and characterization of replication protein from plasmid pGP2 from Acetobacter estunensis  

作　　者：Peter Grones Jozef Grones 

机构地区：[1]不详

出　　处：《Advances in Bioscience and Biotechnology》2010年第5期417-425,共9页生命科学与技术进展（英文）

摘　　要：The Acetobacter estunensis Rep34 protein participates in the replication of bacterial plasmid pGP2. The Rep34 protein of the A. estunensis, was cloned to the expression vector, that ensure fusion with a His-tag sequence (Rep34 His-tagged), over-expressed in Escherichia coli and purified by metal-affinity chromatography to yield a highly purified and active protein. On this purified protein number different activities and motifs were detected. DNA band-shift assays showed that the Rep34 His-tagged protein bound to the regulation region for replication on the linear double-stranded DNA. In the protein was determined phosphatase activity, ATPase activity and protein is possible to unwind double strand DNA.The Acetobacter estunensis Rep34 protein participates in the replication of bacterial plasmid pGP2. The Rep34 protein of the A. estunensis, was cloned to the expression vector, that ensure fusion with a His-tag sequence (Rep34 His-tagged), over-expressed in Escherichia coli and purified by metal-affinity chromatography to yield a highly purified and active protein. On this purified protein number different activities and motifs were detected. DNA band-shift assays showed that the Rep34 His-tagged protein bound to the regulation region for replication on the linear double-stranded DNA. In the protein was determined phosphatase activity, ATPase activity and protein is possible to unwind double strand DNA.

关 键 词：ACETOBACTER Estunensis Rep34 Protein DNA-BINDING ACTIVITY ATPase ACTIVITY PHOSPHATASE ACTIVITY UNWINDING ACTIVITY 

分 类 号：R73[医药卫生—肿瘤]