title: Specific identification of (R)-3-hydroxyacyl-ACP: CoA transacylase gene from Pseudomonas and Burkholderia strains by polymerase chain reaction.
abstract: Polyhydroxyalkanoates (PHA) were biodegradable thermoplastics. Due to their broad applications, direct biosynthesis of PHA from inexpensive substrates, such as carbohydrates, is actively pursued. It has been recently revealed that (R)-3-hydroxyacyl-ACP: CoA transacylase (PhaG) played an important role in this pathway. In this study, a polymerase chain reaction (PCR) protocol was developed for the rapid and specific identification of phaG gene from various bacteria. Using the PCR strategy, the complete open reading frames of two phaG genes from Pseudomonas stutzeri 1317 and Pseudomonas nitroreducens 0802 were cloned from the genomic DNA and functionally expressed in Pseudomonas putida PHAGN-21. Furthermore, this strategy was successful applied in non-Pseudomonas strains, such as Burkholderia. These results suggest that PhaG-mediated pathway of medium-chain-length polyhydroxyalkanoates was widespread among bacteria.
authors.affiliation: MOE Laortoy of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua Unieirsty, Beiing 100084, China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zheng
authors.forename: Jin-Chun
authors.initials: JC
authors.lastname: Chen
authors.forename: Hong-Lei
authors.initials: HL
authors.lastname: Tian
authors.forename: Feng-Feng
authors.initials: FF
authors.lastname: Bei
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Chen
mesh_terms: D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D019117:Burkholderia / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D054813:Polyhydroxyalkanoates / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment
chemical_list: D054813:Polyhydroxyalkanoates; D000217:Acyltransferases; C114682:3-hydroxyacyl-CoA-acyl carrier protein transferase