PMID:21477323

Citation	Wang, ZH, Ma, P, Chen, J, Zhang, J, Chen, CB and Chen, GQ (2011) A transferable heterogeneous two-hybrid system in Escherichia coli based on polyhydroxyalkanoates synthesis regulatory protein PhaR. Microb. Cell Fact. 10:21
Abstract	Polyhydroxyalkanoate (PHA) synthesis regulatory protein PhaR contains a DNA binding domain (DBD) and a PHA granule binding domain (GBD), it anchors to the promoter region of PHA granule-associated protein (PhaP) to repress phaP expression. However, PhaR will bind to PHB granules and be released from phaP promoter region when PHA granules are formed in vivo, initiating expression of phaP gene. Based on this regulatory mechanism, a bacterial two-hybrid system was developed: PhaR was separated into two parts: DBD was used to fuse with the bait, GBD with the prey, and phaP was replaced by a reporter gene lacZ. However, GBD protein expressed in vivo formed inclusion bodies. Thus, PhaP with strong binding ability to PHB granules was employed to replace GBD.
Links	PubMed PMC3079617 Online version:10.1186/1475-2859-10-21
Keywords	Escherichia coli/chemistry; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Polyhydroxyalkanoates/metabolism; Promoter Regions, Genetic; Protein Binding; Protein Structure, Tertiary; Two-Hybrid System Techniques
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