Species identification of Crassostrea and Ostrea oysters by polymerase chain reaction amplification of the 5S rRNA gene.
A specific multiplex polymerase chain reaction (PCR) was developed for the identification of Crassostrea angulata, C. gigas, Ostrea edulis, and O. stentina oyster species. Universal primers were used for the amplification of complete repetition units of 5S rDNA in each of the 4 species. The alignment of the obtained sequences was the basis for the specific design of species-specific primers (ED1, ED2, ST1, ST2, CR1, and CR2) located in the nontranscribed spacer regions. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed identification of Crassostrea and Ostrea species. A multiplex PCR with a set of the 6 designed primers showed that they did not interfere with each other and bound specifically to the DNA target. This genetic marker can be very useful for traceability of the species, application in the management of oyster cultures, and conservation of the genetic resources of the species.
Cross
Ismael
I
Universide Cádiz, Facultad de Ciencias Mar y Ambientales, Laboratorio de Genética, Poligono del Rio San Pedro, 11510 Puerto Real, Cádiz, Spain.
Rebordinos
Laureana
L
Diaz
Edgardo
E
D000818:Animals; D001483:Base Sequence; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D003001:Cloning, Molecular; D049896:Crassostrea / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D021903:DNA, Ribosomal Spacer; D004586:Electrophoresis; D004587:Electrophoresis, Agar Gel / Q000379:methods; D008969:Molecular Sequence Data; D049895:Ostrea / Q000235:genetics*; D010125:Ostreidae; D016133:Polymerase Chain Reaction / Q000379:methods*; D012341:RNA, Ribosomal, 5S / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D017931:DNA Primers; D021903:DNA, Ribosomal Spacer; D012341:RNA, Ribosomal, 5S; D004247:DNA