PCR cloning of type II polyhydroxyalkanoate biosynthesis genes from two Pseudomonas strains.
Two polyhydroxyalkanoate synthase genes, phaC1 from Pseudomonas pseudoalcaligenes HBQ06 and phaC2 from Pseudomonas nitroreducens 0802, were cloned using a PCR cloning strategy based on the type II pha loci property of Pseudomonas strains. The complete open reading frames (ORFs) of phaC1 (P. nitroreducens HBQ06) and phaC2 (P. nitroreducens 0802) were identified from the PCR products. Using the sequence information, the complete PHA synthase genes were PCR cloned directly from the genomic DNA and expressed in Escherichia coli as confirmed by Fourier transform-infrared spectroscopy and gas chromatography. The differences between PhaC1 and PhaC2 were analyzed and the two proteins were suggested to contain different functions and evolution history.
Zhang
G
G
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, PR China.
Hang
X
X
Green
P
P
Ho
K P
KP
Chen
G Q
GQ
D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D013045:Species Specificity
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase; C067032:poly(3-hydroxyalkenoate)polymerase