The actin multigene family and livestock speciation using the polymerase chain reaction

Abstract

Actins constitute a family of highly-conserved multifunctional intracellular proteins, best known as myofibrillar components in striated muscle fibres. Most vertebrate genomes contain numerous actin genes with high sequence homology in protein coding regions but considerable variability in intron number and sizes. This genetic diversity can be utilised for livestock speciation purposes. The high sequence conservation has enabled a single pair of oligonucleotides to be used to prime the polymerase chain reaction (PCR) with DNA extracted from all animals so far studied. Multiple amplification products were obtained which on gel electrophoresis constituted characteristic species-specific 'fingerprints'. The patterns were reproducible, did not vary between individuals of the same breed or between different breeds within a species, and could be generated even from heat-processed muscle held at 120 degrees C for one hour. Given the capacity of PCR to amplify relatively short sequences in highly-degraded DNA, this approach may be suitable for authentication of processed meat products.

Qualitative PCR for the detection of chicken and pork adulteration in goose and mule duck foie gras

“…Actins constitute a multigene family of highly conserved proteins found in all eucaryotic cells 13 and have been studied in a variety of organisms. 14,15 Nucleotide sequences of actin genes are available in the GenBank/EMBL databank for several species. Some of them were used by Watabe et al 16 to design primers act2 and act4 that amplify a DNA fragment of 521 bp in carp and goldfish, which covered 173 amino acids near the C terminus.…”

Species identification in salted products of red snappers by semi-nested PCR–RFLP based on the mitochondrial 12S rRNA gene sequence

“…2). Since the shorter sequence was easy to be amplified from the degraded DNA (Fairborther, Hopwood, Lockley, & Bardsley, 1998;Pardo & Villareal, 2004), primer L231 was designed with H231 to amplify the mtDNA fragment of only about 450 bp in the second step PCR.…”

Species identification in salted products of red snappers by semi-nested PCR-RFLP based on the mitochondrial 12S rRNA gene sequence

“…2). Since the shorter sequence was easy to be amplified from the degraded DNA (Fairborther, Hopwood, Lockley, & Bardsley, 1998;Pardo & Villareal, 2004), primer L 231 was designed with H 231 to Although the high degree of DNA sequence homology (95-98%, depending on the species being compared) in RDM459s was observed, four available restriction enzyme sites were selected and species authentication could be fulfilled by the comparison of RFLP profiles. The method of semi-nested PCR-RFLP presented here could extend to the species identification of more salted fish products.…”