Prospecting for novel lipase genes using PCR.
A PCR method suitable for the isolation of lipase genes directly from environmental DNA is described. The problems associated with the low levels of similarity between lipase genes were overcome by extensive analysis of conserved regions and careful primer design. Using this method, a lipase gene (oli-lipase) was isolated directly from environmental DNA. This lipase showed less than 20% similarity with other known lipases at the amino acid level. The study also revealed that distantly related members of the alpha/beta hydrolase superfamily share similar conserved motifs with the lipases, thus making these genes targets for gene prospecting by PCR.
Bell
Philip J L
PJL
Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia1.
Sunna
Anwar
A
Physikalische Biochemie, Universitat Potsdam, Karl-Liebknecht-Strasse 24-25 D-14476 Golm, Germany2.
Gibbs
Moreland D
MD
Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia1.
Curach
Natalie C
NC
Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia1.
Nevalainen
Helena
H
Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia1.
Bergquist
Peter L
PL
Division of Molecular Medicine, University of Auckland School of Medicine, Private Bag 92109, Auckland, New Zealand3.|Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia1.
D001407:Bacillus / Q000235:genetics; D018533:Biomass; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial; D004777:Environment; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D004269:DNA, Bacterial; D006867:Hydrolases; D008049:Lipase