Reliable amplification of actin genes facilitates deep-level phylogeny.
The gene for actin as a highly conserved and functionally essential genetic element is developing into a major tool for phylogenetic analysis within a broad organismic range. We therefore propose a set of universally applicable primers that allow reliable amplification of actin genes. For primer construction the amino acid sequences of 57 actin genes comprising fungi, animals, plants and protists were analysed, aligned and used for the definition of six well-conserved regions which are suitable as priming sites in PCR amplification experiments. Ten primers were designed for specific in vitro amplification of actin gene fragments from a wide range of microorganisms. The corresponding gene fragments provide a strong basis to isolate nearly complete actin genes for further molecular characterization and for establishing phylogenies based on actin gene trees.
Voigt
K
K
Fungal Reference Centre and Department of General Microbiology and Microbial Genetics, Friedrich-Schiller-Universität Jena, Institut für Mikrobiologie, Germany. b5kevo@rz.uni-jena.de
Wöstemeyer
J
J
D020095:Absidia / Q000235:genetics; D000199:Actins / Q000235:genetics*; D000595:Amino Acid Sequence; D017124:Conserved Sequence; D017931:DNA Primers; D019143:Evolution, Molecular; D005658:Fungi / Q000145:classification*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010802:Phylogeny; D016415:Sequence Alignment
D000199:Actins; D017931:DNA Primers