• Title/Summary/Keyword: Cytochrome b gene

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Development of PCR Assay for Identification of Buffalo Meat

  • Rajapaksha, W.R.A.K.J.S.;Thilakaratne, I.D.S.I.P.;Chandrasiri, A.D.N.;Niroshan, T.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1046-1048
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    • 2003
  • A polymerase chain reaction (PCR) assay was developed to differentiate buffalo meat from the meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon sambhur (Cervus unicolor unicolor), cattle (Bovine), goat (Caprine), pig (Porcine), and sheep (Ovine). A set of primers were designed according to the sequence of the mitochondrial cytochrome b gene of bubalus bubalis and by PCR amplification a band of approximately 242 bp band was observed with buffalo DNA. These primers did not cross-react with DNA of other animal species tested in the study under the specified reaction conditions. A band of 649 bp was observed for all animal species tested when DNA was amplified with the universal primers indicating the presence of mitochondrial DNA in the samples. The technique was sensitive enough to identify rotten (10 days post slaughter), dried and cooked buffalo meat. The absence of a cross reaction with human DNA using the buffalo specific primers eliminates possible false positive reactions.

Adulteration of Caprine Milk Products by Bovine Milk in Korea (국내 시판 산양유제품 내 젖소 유성분의 혼입에 대한 조사 연구)

  • Jung, Tae-Hwan;Jeon, Woo-Min;Han, Kyoung-Sik
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.127-131
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    • 2013
  • The aim of this study was to investigate adulteration of caprine milk products by bovine milk using biomolecular techniques with bovine-specific primers for the mitochondrial cytochrome b gene. Polymerase chain reaction (PCR) and real-time PCR assays were applied to caprine milk products including infant formula, city milk, and fermented milk. The results indicated that six out of the eight caprine infant formula products tested contained bovine milk components. In addition, two of the three tested caprine city milk products and two caprine fermented milk products were shown to be adulterated with bovine milk. Conventional PCR results corroborated with results obtained by real-time PCR. This study demonstrates that DNA-based species identification procedures would be useful and applicable in routine examinations of the dairy industry to ensure the quality and safety of dairy foods.

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DNA Barcoding of Antarctic Freshwater Copepod Boeckella poppei (Crustacea: Copepoda: Calanoida: Centropagidae) Inhabiting King George Island, South Shetland Islands, Antarctica

  • Kang, Seunghyun;Jo, Euna
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.4
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    • pp.396-399
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    • 2020
  • The Antarctic freshwater copepod, Boeckella poppei (Mrazek, 1901), has the widest range of distribution extending from southern South America to Antarctic continent, among all Boeckella species. Boeckella poppei is the only freshwater copepod known to be inhabiting the Antarctic continent. In present study, we analyzed the DNA barcodes of the mitochondrial cytochrome c oxidase subunit I (COI) gene of B. poppei from King George Island, Antarctica. The intraspecific genetic distances varied from 0% to 13% and interspecific genetic distances ranged from 11% to 14%. The overlap of DNA barcode gap suggests careful threshold-based delimitation of species boundaries.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

Development and validation of a PCR method to discriminate between Branchiostegus japonicus and Branchiostegus albus (옥돔과 옥두어 판별을 위한 PCR 검사법 개발과 검증)

  • Kim, Na-Ye-Seul;Yang, Ji-Young;Kim, Jung-Beom
    • Korean Journal of Food Science and Technology
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    • v.51 no.3
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    • pp.295-299
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    • 2019
  • We developed and validated species-specific primers for Branchiostegus japonicus and Branchiostegus albus to prevent the sale of B. albus as B. japonicus. Primers for B. japonicus and B. albus were designed against the cytochrome b gene. Multiplex PCR showed a 288 bp amplicon for B. japonicus, a 159 bp amplicon for B. albus, and a 502 bp amplicon for the internal control. The PCR product bands for B. japonicus, B. albus, and the internal control were present at 1 ng each. The specificity and sensitivity of the primers developed in this study were validated by testing 38 B. japonicus strains and 13 B. albus strains. Using this monitoring method, fake fish did not appear due to the agreement between the experimental results and the species. Therefore, the developed multiplex PCR method was suitable for differentiating B. japonicus and B. albus.

Discordance between Morphological and Molecular Variations of the Genus Macroramphosus (Macroramphosidae) from Korea (한국산 대주둥치속(대주둥치과) 어류의 형태와 분자 변이의 불일치)

  • Sohn, Min-Soo;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.199-209
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    • 2020
  • In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.

Genetic Species Identification by Sequencing Analysis of Nuclear and Mitochondrial Genes for Albino Misgurnus Species from Korea (우리나라 미꾸리속(genus Misgurnus) 알비노 개체의 미토콘드리아 및 핵 유전자 염기서열 분석에 의한 유전적 동정)

  • Song, Ha-Youn;Moon, Shin-Joo;Kim, Keun-Sik;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.29 no.2
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    • pp.139-145
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    • 2017
  • The spontaneous color mutant, albino individuals of genus Misgurnus, are rarely discovered in Korea and there are difficult to identify morphological species due to lack melanin pigmentation. In this study, we developed a genetic identification method for the species of albino Misgurnus individuals based on phylogenetic analysis by using recombination activating gene 1 (rag1) and cytochrome b (cytb) region of mitochondrial DNA. As a result of molecular phylogenetic analysis, three clades were identified as Misgurnus mizolepis, M. anguillicaudatus and M. mohoity. The homology of the cytb sequences of M. mohoity was best match to that of M. mohoity sequences in GenBank database. As a result of species identification of 25 albino Misgurnus individuals based on the phylogenetic tree, the red-eye type was identified as 16 M. anguillicaudatus and one M. mizolepis. The remaining three individuals were identified as one M. mizolepis ♀${\times}$M. anguillicaudatus ♂, and two M. mohoity ♀${\times}$M. anguillicaudatus ♂, respectively. In addition, the five black-eye type individuals were identified as one M. anguillicaudatus, three M. mizolepis and one M. mohoity. Therefore, this genetic identification method will be an useful techniques for species or hybrid identification in genus Misgurnus.

Genetic origin identification of Siberian chipmunks (Tamias sibiricus) in pet shops of South Korea

  • Lee, Seo-Jin;Jung, Gil-A;Min, Mi-Sook;Kim, Chuel-Kyu;Lee, Hang;Kim, Chang-Bae;Lee, Mu-Yeong
    • Animal cells and systems
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    • v.15 no.2
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    • pp.161-168
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    • 2011
  • Siberian chipmunks, Tamias sibiricus, are one of several popular companion animals found in the pet shops of South Korea. At present, however, there have been no studies done in South Korea examining their origin even though they could be potential carriers of zoonotic diseases, and are a species of concern for efficient conservation and management strategies. Sequences of the mitochondrial cytochrome b gene (1140 bp) were determined to investigate the origin of Siberian chipmunks sold in four South Korean pet shops through comparison with sequence data from animals of known locality. Nine Siberian chipmunks were collected from pet shops in South Korea, which resulted in nine haplotypes. One (AR) of these coincided with the haplotype previously described. Phylogenetic and network analyses using 53 haplotypes including 45 haplotypes from GenBank showed three phylogenetic groups in South Korea, almost concordant to locality, designated as northern, central, and southern parts as described in a previous study. Of the nine individuals examined from the pet shops, eight were clustered into the northern phylogroup but one (cgrb9153) was grouped with the southern phylogroup, implying that at least the Siberian chipmunks examined in this study did not originate from other countries. It is likely that most individuals sold in the pet shops of Seoul were caught in the wild in Gyeonggi-do and Gangwon-do, or are maternal descendants of captive-bred individuals originating from the northern part of South Korea. It is recommended that conservation and management units of Korean chipmunks should be examined in further detail.

Inhibitory effects of heavy metals on CYP1A expression in eel hepatocyte cultures (뱀장어 배양 간세포에서의 Cytochrome P4501A (CYP1A) 유전자 발현에 대한 중금속들의 억제효과)

  • Kwon, Hyuk-Chu;Maeng, Joon-Ho;Choi, Seong-Hee
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.245-254
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    • 2010
  • Effects of heavy metal ions on the gene expression of cytochrome P4501A (CYP1A) were examined in cultured eel hepatocytes. When the expression of CYP1A mRNA was measured by RT-PCR after incubation of eel hepatocytes with benzo[$\alpha$]pyrene (B[$\alpha$]P) at concentrations of 10-8~10-5 M, the CYP1A expression increased with B[$\alpha$]P treatment in a dose dependent manner, showing significant increase at concentrations more than 10-7 M. When the eel hepatocyte was treated with cadmium (10-6 and 10-5 M), the expression of CYP1A was inhibited and especially at higher concentration (10-5 M). The inhibition of CYP1A expression by cadmium was also observed in cells treated with B[$\alpha$]P. In another study, effects of heavy metal ions on the expression of CYP1A were examined in cultured hepatocytes isolated from eel which was treated previously with B[$\alpha$]P in vivo. Hepatocytes isolated from the liver of eel taken at 48 hours after injection of B[$\alpha$]P (10 mg/kg) were cultured for 2 days with cadmium, copper, lead or zinc (10-6 and 10-5 M). The expression of CYP1A was found to be suppressed by the metal ions compared with the control in which CYP1A was induced with previous treatment of B[$\alpha$]P in vivo. The present results may provide an important basic information for studying the effects of heavy metal ions on CYP1A expression in other species of fish and studying toxicological mechanisms of heavy metal ions in aquatic livings.