• Title/Summary/Keyword: Specific primer

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Infestation Status of Clonorchis sinensis Metacercariae in Cyprinid Fish from four Artificial Lakes in South Korea (소양호, 남양호, 안동호 및 춘천호에 서식하는 잉어과 어류의 간흡충 Clonorchis sinensis 감염 조사)

  • Jeon, Sang Woo;Hur, Jun Wook;Song, Jun Young;Jee, Bo Young;Lee, Jeong Ho;Kwon, Joon Yeong;Kwon, Se Ryun
    • Journal of Marine Life Science
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    • v.3 no.2
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    • pp.96-100
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    • 2018
  • Clonorchis sinensis is a main parasite that infects humans by making freshwater fish as an intermediate host in South Korea. There are so many reports about the infestation status of Clonorchis sinensis metacercariae (CsMc) in freshwater fish living in the river, but there are a few studies of fish in the lake. In this study, we examined CsMc in Cyprinid fish sampled from Soyang Lake, Namyang Lake, Andong Lake and Chunchun Lake from 2016 to 2017. Metacercaria of trematodes were found from Hemiculter eigenmanni and Carassius auratus in Namyang Lake, and Zacco platypus and Opsarichthys uncirostris in Soyang Lake. As a result of PCR using Clonorchis sinensis specific primer sets, it was confirmed that the metacercariae from Hemiculter eigenmanni in Namyang Lake was CsMc. This study provides information on CsMc infestation status of Cyprinid fish in four lakes and it is the first report of CsMc infestation in Namyang Lake.

Effects of the Proliferation of Beneficial and Harmful Enteric Bacteria after Intake of Soybean Fermentation (Zen) Produced by a Mixture of Lactobacilli and Saccharomyces (Lactobacilli와 Saccharomyces 혼합균주의 대두발효액(Zen) 섭취 후 장내 유익세균과 유해세균의 증식에 미친 영향)

  • Won, Ryu Seo;Lee, Hyung H.
    • Journal of Naturopathy
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    • v.8 no.1
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    • pp.1-10
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    • 2019
  • Purpose: The purpose of this study was to investigate the increase or decrease of important intestinal beneficial bacteria and inhibitory bacteria in 30 stools of clinical subjects after ingesting Zen fermentation broth as a mixed microbial fermentation solution for eight weeks. Methods: Intestinal bacteria were identified by PCR amplification using specific primers. Results: Bifidobacterium genus gi% of test group ingested Zen-fermented broth was 55.15% before and 70.1% after ingestion, so it was a significant difference (p<.009). Lactobacillus genus of the test group was 46.87% before and 60.91% after ingestion, it was a significant difference (p<.01). Clostridium genus of the test group was 85.64% before and 65.99% after ingestion. There was a significant difference (p<.017) as the pre-post-difference decreased to -19.65%. Bacteroides genus of the test group was 17.11% before and 20.22% after ingestion. There was a significant difference (p<.048) as the pre-post-difference increased to 3.11%. Prevotella genus of the test group was 14.01% before and 16.79% after ingestion, so it was not a significant difference. Conclusions: Intestinal bacteria increased the proliferation of beneficial bacteria and suppressed harmful bacteria in the intestines after ingesting the Zen-fermented broth of the mixed microorganism. The Zen fermentation broth evaluated as a beneficial drink for intestinal health.

Distribution of Beneficial Bacteria in the Intestines after Enzamin Ingestion of Bacillus subtilis AK Strain Fermentation (Bacillus subtilis AK균 발효액(Enzamin)의 섭취 후 장내 유익세균의 분포조사)

  • Ryu, Seo Won;Lee, Hyung H.
    • Journal of Naturopathy
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    • v.7 no.2
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    • pp.27-38
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    • 2018
  • Purpose: The purpose of this study was to investigate whether intestinal proliferation is promoted in beneficial intestinal bacteria or decreased in harmful bacteria before and after ingesting Bacillus fermentation broth (ENM) for 8 weeks in the 16 subjects. Method: Intestinal bacteria were identified by PCR amplification using specific 16S rRNA primers. Results: The Bifidobacterium gene index(%)(gi%) increased to 58.92% in the control group and 69.53% in the test group after the ingestion of ENM, but there was no significant difference. Lactobacillus gi% increased significantly (49.37% in the control and 66.43% in the test) (p<.029). Clostridium gi% was significantly decreased after treatment (83.16% in the control and 67.76% in the test) (p<.077). Bacteroides gi% increased significantly (12.58% in the control and 20.87% in the test) after ingesting (p<.095). Prevotella gi% increased significantly (7.55% in the control and 17.28% in the test) after ingesting (p<.005). After ingesting, the median bacteria increased significantly in the control (20.06%) and the test (35.88%) (p<.001). Conclusions: After ingestion of the ENM, the number of beneficial bacteria increased and the number of harmful bacteria Clostridium tended to decrease. This suggests that ingestion of the Bacillus fermented beverage ENM has an effect on the proliferation of intestinal bacteria.

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Characterization of the DGAT1 Gene in the Korean Holstein Dairy Cattle Population (한국 Holstein종 유우집단의 DGAT1 유전자의 특성분석)

  • Son, Ji-Young;Jeong, Hang-Jin;Yu, Seong-Lan;Lee, Jun-Heon;Do, Chang-Hee;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.36 no.2
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    • pp.167-177
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    • 2009
  • This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).

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Sequence and Genetic Variation of Mitochondrial DNA D-loop Region in Korean Cattle (한우 Mitochondrial DNA D-loop 영역의 염기서열 및 유전변이)

  • Chung, E.R.;Kim, W.T.;Kim, Y.S.;Lee, J.K.;Han, S.K.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.181-190
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    • 2002
  • This study was performed to determine sequences of the mt DNA D-loop region, including $tRNA^{Pro}$ and $tRNA^{Pre}$ and to analysis sequence variation polymorphism in Korean cattle. The resulting sequencies were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify an 1142bp between nucleotides 15061 and 404 within the D-loop region of mt DNA using specific primers. Korean cattle showed 24 polymorphic sites by nucleotide substitutions and insertions of single base pairs. About 50% of polymorphic sites were found in positions 16042 to 16122 with the most variable region. Among these polymorphic sites, variations at 16055, 16230 and 16260 bp were detected as new sequence variants in Korean cattle. These specific polymorphic sites have not been reported in the Japanese black cattle and European cattle. Therefore, mt DNA variants in the D-loop region may be used as genetic markers for specifying Korean cattle. The frequencies of positions 169, 16302, 16093, 16042, 16119 with a high level of sequence polymorphism were 0.81, 0.56, 0.56, 0.50 and 0.43, respectively. In comparison of genetic distances, Korean cattle showed the more closely to European cattle as Bos taurus than Bos indicus such as African and India breeds. In conclusion, these mt DNA sequence polymorphisms in the D-loop region for Korean cattle may be useful for the analysis of cytoplasmic genetic variation and associations with economic important traits and genetic analysis of maternal lineage.

Sex Determination of Porcine IVF Embryos by Polymerase Chain Reaction (PCR) (중합효소연쇄반응을 이용한 돼지 체외수정란의 성감별)

  • 강미선;김용준;이해이;유일정
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.203-211
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    • 2001
  • This study was carried out to determine sex of porcine embryos produced by in vitro fertilization. Porcine oocyte-cumulus complexes were cultured in BSA-free North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (10%), cystein (0.1 mg/ml) and hormonal supplement (10 IU eCG and 10 IU hCG per ml) for 20~22 hrs. They were then cultured in the same medium but without hormonal supplement for additional 20~22 hrs. After culture, cumulus cells were removed and oocytes were co-incubated for 6 hrs with four different concentrations (5$\times$10$^4$, 2.5$\times$ 10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) of porcine sperm. After fertilization, oocytes were transferred into NCSU 23 with 0.4% BSA medium. The cleavage and blastocyst formation rates were evaluated at 48 and 144 hrs, respectively. In this study, the polymerase chain reaction (PCR) was used to determine the sex of porcine embryos in the stage of blastocyst. The PCR was performed using a set of oligonucleotide primers (5‘-TCATGGACCAGGTAGGGAAT-3', 5’-GAAAGACACGTCCTTGGA GA-3') for 491 bp fragment of porcine male-specific DNA sequence. In the flour different sperm concentration (5$\times$10$^4$, 2.5$\times$10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) for fertilization condition, the cleavage rate was 55.95, 67.88, 60.18 and 47.60%, respectivety, and the development rate of blastocysts was 16.03, 20.40, 21.41 and 12.37%, respectively. At 5.0$\times$10$^4$and 2.5$\times$10$^{5}$ of sperm concentrations per ml cleavage rate and development rate of blastocyst were higher than those of 5.0$\times$10$^4$and l0$\times$10$^{5}$ of sperm concentration (P<0.01). The male of porcine embryos was detected at 491 bp by PCR, and 18 of the 31 porcine blastocysts were the male (58.1%) and the rest 13 were the female(41.9%).

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Phylogenetic Analysis of the Genus Phellinus by Comparing the Sequences of Internal Transcribed Spacers and 5.8S Ribosomal DNA (Ribosomal DNA의 Internal Transcribed Spacer(ITS) 부위의 염기서열분석에 의한 Phellinus속의 계통분석에 관한 연구)

  • Chung, Ji-Won;Kim, Gi-Young;Ha, Myung-Gui;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.124-131
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    • 1999
  • This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.

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Comparative Assessment of Specific Genes of Bacteria and Enzyme over Water Quality Parameters by Quantitative PCR in Uncontrolled Landfill (정량 PCR을 이용한 비위생 매립지의 특정 세균 및 효소 유전자와 수질인자와의 상관관계 평가)

  • Han, Ji-Sun;Sung, Eun-Hae;Park, Hun-Ju;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.8
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    • pp.895-903
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    • 2007
  • As for the increasing demanding on the development of direct-ecological landfill monitoring methods, it is needed for critically defining the condition of landfills and their influence on the environment, quantifying the amount of enzymes and bacteria mainly concerned with biochemical reaction in the landfills. This study was thus conducted to understand the fates of contaminants in association with groundwater quality parameters. For the study, groundwater was seasonally sampled from four closed unsanitary landfills(i.e. Cheonan(C), Wonju(W), Nonsan(N), Pyeongtaek(P) sites) in which microbial diversity was simultaneously obtained by 16S rDNA methods. Subsequently, a number of primer sets were prepared for quantifying the specific gene of representative bacteria and the gene of encoding enzymes dominantly found in the landfills. The relationship between water quality parameters and gene quantification were compared based on correlation factors. Correlation between DSR(Sulfate reduction bacteria) gene and BOD(Biochemical Oxygen Demand) was greater than 0.8 while NSR(Nitrification bacteria-Nitrospira sp.) gene and nitrate were related more than 0.9. A stabilization indicator(BOD/COD) and MTOT(Methane Oxidation bacteria), MCR(Methyl coenzyme M reductase), Dde(Dechloromonas denitrificans) genes were correlated over 0.8, but ferric iron and Fli(Ferribacterium limineticm) gene were at the lowest of 0.7. For MTOT, it was at the highest related at 100% over BOD/COD. In addition, anaerobic genes(i.e., nirS-Nitrite reductase, MCR. Dde, DSR) and DO were also related more than 0.8, which showing anaerobic reactions generally dependant upon DO. As demonstrated in the study, molecular biological investigation and water quality parameters are highly co-linked, so that quantitative real-time PCR could be cooperatively used for assessing landfill stabilization in association with the conventional monitoring parameters.

A Comparison of Gene Extraction Methods for the Identification of Raw Materials from Processed Meat Products (식육추출가공품의 사용원료 확인을 위한 유전자추출 방법의 비교 및 검토)

  • Park, Yong-Chjun;Kim, Mi-Ra;Lim, Ji-Young;Park, Young-Eun;Shin, Jun-Ho;Hwang, Cho-Rong;Lim, Jan-Di;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.146-151
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    • 2012
  • In this study, effective gene extraction methods were compared to identify raw materials of processed meat products through molecular biological methods. Species specific primers were used to identify ingredients of processed foods and, as a sample, 13 kinds of processed meat products including beef, pork and chicken. According to the type of sample, 13 kinds of samples were classified into liquid type, source type and powder type. The samples were pre-treated (centrifugation) and (or) performed Whole Gene Amplification (WGA) kit for amplification of the extracted DNA. As a result, it was possible to identify the raw material of products through the centrifugation of sample 1 ml for liquid type of processed meat products. For source type of products after gene extraction, it was required to perform WGA for the identification of ingredients. For powder type products did not required any further pre-treatment and WGA. In this study, it was an opportunity to confirm the possibility of identification of raw material from the gene extraction of processed meat products and this method could be used to examine the authenticity of raw material of products.

Species-specific Expression of Rpia Transcript in Cumulus-oocyte-complex (난자-난구세포 복합체에서 발현하는 Rpia 유전자의 종 특이적 발현)

  • Kim, Yun-Sun;Yoon, Se-Jin;Kim, Eun-Young;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.95-106
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    • 2007
  • Objective: We previously identified differentially expressed genes (DEGs) between germinal vesicle (GV) and metaphase II (MII) mouse oocyte. The present study was accomplished as a preliminary study to elucidate the role of ribose 5-phosphate isomerase A (Rpia), the essential enzyme of the pentose phosphate pathway (PPP), in oocyte maturation. We observed expression of Rpia in the mouse and porcine oocytes. Methods: Expression pattern of the 11 MII-selective DEGs in various tissues was evaluated using RT-PCR and selected 4 genes highly expressed in the ovary. According to the oocyte-selective expression profile, we selected Rpia as a target for this study. We identified the porcine Rpia sequence using EST clustering technique, since it is not yet registered in public databases. Results: The extended porcine Rpia nucleotide sequence was submitted and registered to GenBank (accession number EF213106). We prepared primers for porcine Rpia according to this sequence. In contrast to the oocyte-specific expression in the mouse, Rpia was expressed in porcine cumulus and granulosa cells as well as in oocytes. Conclusion: This is the first report on the characterization of the Rpia gene in the mouse and porcine ovarian cells. Results of the present study suggest that the mouse and porcine COCs employ different mechanism of glucose metabolism. Therefore, the different metabolic pathways during in vitro oocyte maturation (IVM) in different species may lead different maturation rates. It is required to study further regarding the role of Rpia in glucose metabolism of oocytes and follicular cell fore exploring the regulatory mechanism of oocyte maturation as well as for finding the finest culture conditions for in vitro maturation.