G. Yoshizaki et al., CLONING AND CHARACTERIZATION OF PEJERREY MITOCHONDRIAL-DNA AND ITS APPLICATION FOR RFLP ANALYSIS, Journal of Fish Biology, 51(1), 1997, pp. 193-203
Probes were cloned, characterized, and developed for all regions of th
e mitochondrial DNA (mtDNA) of pejerrey Odontesthes bonariensis to pro
vide the basis for the study of genetic diversity of South American at
herinopsinii and to enable species identification from small amounts o
f tissue. The mtDNA was extracted from liver and cleaved with Eco RI,
producing four fragments (7.4, 3.4, 3.1 and 2.9 kb) which were cloned
using pUC118 plasmid vectors. Sequence analysis from both ends of the
fragments showed that they encode tRNA (Asp, Phe, and Ser-TGA), 12 S r
RNA, cytochrome oxidase (GO) II, NADH 4, 5, and 6, and the D-loop, and
that the relative positions of these genes are identical to those in
the mtDNA of other teleosts. A comparison of homology with carp mtDNA
nucleotide sequences revealed that tRNA (Phe and Ser-TGA) and CO II we
re relatively conserved, whereas the D-loop region was highly divergen
t. The cloned mtDNA probes detected mtDNA fragments from about 800 ng
of total DNA extracted from liver, muscle, and single embryos of O. bo
nariensis, and were effective for restriction length fragment polymorp
hism (RFLP) analysis of Patagonina hatcheri, the most distant atherino
psine relative of pejerrey. The cloned mtDNA probes may be useful for
the analysis of genetic diversity and non-destructive species identifi
cation, including the examination of eggs, larvae and juveniles. The m
tDNA sequences reported here provide the basis for the design of prime
rs for PCR-based RFLP analysis. (C) 1997 The Fisheries Society of the
British Isles.