• Journal of Life Science
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• v.21 no.3
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• pp.341-348
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• 2011

In an effort to gain greater understanding of the maternal lineages of the Jeju native pig (JNP), we analyzed the mitochondrial DNA (mtDNA) CYTB gene and compared it with those of other pig breeds. PCR-RFLP analysis was conducted with six pig breeds including JNP, and then the RFLP patterns allowed for the separation of the pig breeds into two distinct haplotypes (mtCYTB1 and mtCYTB2). The JNP CYTB sequences were detected in both the European and Asian breed clusters on the phylogenetic tree. The J2 group was sorted with the indigenous cluster of Asian pig lineages and was related closely to Chinese native pig breeds, but a second group, J1, was sorted with the European pig lineages and appeared to be related to Spanish Iberian native pigs, rather than to Asian breeds. These results indicate that the JNP currently raised on Jeju Island have two major maternal origins estimated in Asian and European pigs. We concluded that the JNP that share a common lineage with indigenous Asian pigs were domesticated in the distant past, originating from pigs that were already being raised elsewhere at that time, and that the European pig breeds introduced in the twentieth century have also contributed to the formation of this pig population.

• Analytical Science and Technology
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• v.18 no.3
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• pp.257-262
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• 2005

Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

• Animal Systematics, Evolution and Diversity
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• v.36 no.3
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• pp.216-221
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• 2020

Parrots are common targets for illegal trade because of their beauty and high price. Accurate identification is necessary for the prevention of illegal trade and conservation of parrots. In the present study, mitochondrial markers of cytochrome b (CYTB) gene were used to identify parrot species from Korean zoos. Totally, 27 samples were collected from Seoul Zoo, Cheongju Zoo, and Uchi Zoo. After collection, total DNA of samples was extracted and used for PCR amplification. CYTB fragments were sequenced from all samples examined. The obtained sequences were used for GenBank blast, distance estimation, and phylogenetic analysis. All species were identified using CYTB sequences that determined 27 samples belong to 13 species in 7 genera, and 3 families. Our finding demonstrated the usefulness of CYTB sequences for identifying parrot species in Korean zoos.

• Parasites, Hosts and Diseases
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• v.57 no.5
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• pp.481-487
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• 2019

Mitochondrial DNA sequence variability of Spirometra erinaceieuropaei in GenBank was observed by reinvestigation of mitochondrial cox1 and cytb sequences. The DNA sequences were analyzed in this study, comprising complete DNA sequences of cox1 (n=239) and cytb (n=213) genes. The 10 complete mitochondrial DNA sequences of Spirometra species were compared with those of Korea, China and Japan. The sequences were analyzed for nucleotide composition, conserved sites, variable sites, singleton sites and parsimony-informative sites. Phylogenetic analyses was done using neighbor joining, maximum parsimony, Bayesian inference and maximum-likelihood on cox1 and cytb sequences of Spirometra species. These polymorphic sites identified 148 (cox1) and 83 (cytb) haplotypes within 239 and 213 isolates from 3 Asian countries. Phylogenetic tree topologies were presented high-level confidence values for the 2 major branches of 2 Spirometra species containing S. erinaceieuropaei and S. decipiens, and S. decipiens sub-clades including all sequences registered as S. erinaceieuropaei in cox1 and cytb genes. These results indicated that mitochondrial haplotypes of S. erinaceieuropaei and S. decipiens were found in the 3 Asian countries.

• Journal of Life Science
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• v.30 no.12
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• pp.1063-1069
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• 2020

Two crucian carp species (Carassius auratus and C. cuvieri) inhabit Lake Yedang in South Korea, and C. auratus is known to be native to Korea. Classification of these two freshwater fish species is often confused because of their morphological similarity. To distinguish the two species, we conducted phylogenetic and population genetic analyses of C. auratus and C. cuvieri based on their mitochondrial DNA sequences of the cytochrome b gene (CYTB). We also compared our partial CYTB sequence (<1,056 bp) with 10 Chinese, nine Japanese, and two Russian crucian carp fishes. The results of our phylogenetic analysis showed that C. auratus and C. cuvieri were clearly divided into two phylogroups. The nucleotide diversity (π) of C. auratus from Korea, China, and Japan showed a range of 0.146%~0.421%, while the range of π of C. cuvieri from Korea and Japan was lower than those of C. auratus (0.0%~0.054%). Moreover, the comparison of CYTB divergence among crucian carp fishes in China, Japan, and Korea indicated that Korean Carassius fishes were distantly related to those from China and Japan, with two exceptions: the pairwise Fst value between Korean C. auratus and northern Chinese C. auratus was not significantly different. In addition, no significant genetic divergence between Korean and Japanese C. cuvieri was detected. We conclude that, despite the morphological similarities, C. auratus and C. cuvieri should be considered as separate freshwater fish resources in conservation efforts for genetic diversity.

• Animal cells and systems
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• v.11 no.2
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• pp.165-168
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• 2007

The nucleotide sequences of a male-specific marker sex determining region Y (SRY) gene and a mitochondrial cytochrome B (CYTB) gene were characterized and analyzed to establish a molecular method for identification of species and sex of Korean roe deer (Capreolus pygargus tianschanicus). Similarity search result of SRY sequences showed very similar result to those reported in Moose (Alces alces) and Reindeer (Rangifer tarandus), both of which had 95.9% similarity in identity. CYTB genes were very similar to those reported in Siberian roe deer (C. pygargus pygargus) which had 98.6% similarity and not to European roe deer (C. capreolus), suggesting that the DNA samples tested were of Siberian roe deer lineage. Polymerase chain reaction (PCR)-based sex typing successfully discriminated between carcasses of male and female roe deer. Males had SRY band on agarose gels and females did not. The result of this molecular sex typing provided similar information with that obtained by genital organ observation. Therefore, this molecular method using male specific marker SRY and mitochondrial CYTB genes would be very useful for identification of the species and sex of the carcass remains of roe deer.

• Korean Journal of Ichthyology
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• v.34 no.3
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• pp.208-217
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• 2022

We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

• Fisheries and Aquatic Sciences
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• v.27 no.1
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• pp.48-55
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• 2024

The study assessed the utility of mitochondrial DNA for identifying a deceased rorqual discovered off the western coast of India. Both the COI and Cytb genes exhibited remarkable 99-100% similarity with the GenBank sequence of Balaenoptera musculus through a global BLAST search, confirming their affiliation with this species. Inter-species genetic distances for COI and Cytb genes ranged from 6.75% to 9.80% and 7.37% to 10.96% respectively, compared with other Balaenopteridae species. The Bayesian phylogenies constructed based on both COI and Cytb genes demonstrated clear and separate clustering for all Balaenopteridae species, further reaffirming their distinctiveness, while concurrently revealing a cohesive clustering pattern of the generated sequences within the B. musculus clade. Beyond species confirmation, this study provides valuable insights into the presence of live and deceased B. musculus individuals within Indian marine ecosystems. This information holds significant potential for guiding conservation efforts aimed at safeguarding Important Marine Mammal Areas (IMMAs) in India over the long term.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.355-360
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• 2013

Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

• Korean Journal of Ichthyology
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• v.36 no.2
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• pp.139-155
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• 2024

Using the cytb gene region of the mitochondrial DNA of eight populations of Sarcocheilichthys nigripinnis morii and five populations of S. variegatus wakiyae, which belong to the genus Sarcocheilichthys from Korea, the genetic diversity and molecular phylogenetic relationships of each population were examined. As a result of the analysis, it was confirmed that the S. variegatus wakiyae population had higher genetic diversity than the S. nigripinnis morii population. In the phylogenetic tree of genus Sarcocheilichthys fish in Korea based on the cytb gene, the Yeongsan River (YSR) population of S. variegatus wakiyae forms a clade with the Tamjin River (TJR), Yeongsan River (YSR), and Seomjin River (SJR) population of S. nigripinnis morii, and genetic relationships that do not align with the current classification system were observed. Meanwhile, on the nuclear DNA phylogenetic tree, S. variegatus wakiyae and S. nigripinnis morii could be clearly distinguished, showing mitonuclear inconsistency where mitochondrial and nuclear DNA conflicted on the phylogenetic tree. The Seomjin River (SJR) population of S. nigripinnis morii was translocated to the Dongjin River (DJR) population, haplotype from which crossbreeding was presumed to have occurred was confirmed. Among the rivers flowing into the East Sea, the S. nigripinnis morii population is known to have been introduced and inhabit only the Hyeongsan River (HSR), and it is presumed to be a population formed by translocation from the Han River (HR) population, with a haplotype representing a unique genetic group also confirmed. The Han River (HR), Geum River (GR), and Mangyeong River (MGR) populations of S. nigripinnis morii formed a genetically identical population with S. czerskii and S. soldatovi distributed north of the Yalu River, and accordingly, a taxonomic reexamination was required through morphological and molecular phylogenetic studies by securing various specimens.