• Title/Summary/Keyword: TaqI

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Association Between MSTN Gene Polymorphism and Growth Traits in Landrace Pigs (돼지 Landrace 품종에서 Myostatin 유전자의 유전적 다형성과 성장형질과의 연관성)

  • Cho, I.C.;Choi, Y.L.;Ko, M.S.;Kim, H.S.;Lee, J.G.;Jeon, J.T.;Han, Sang-Hyun
    • Journal of Animal Science and Technology
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    • v.47 no.2
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    • pp.159-166
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    • 2005
  • Porcine myostatin(MS1N) gene plays a key role in the differentiation of myoblast and muscle development. Genetic polymorphism was screened by single stranded conformation polymorphism(SSCP) analysis and subsequent DNA sequencing detected a nucleotide substitution(C2150T) in exon 3 of MSIN gene. Phenotypic association of the polymorphism was tested in a Landrace population and positive effects of the allele T for lean growth traits were found in the population. Even though it is not significant, the pigs have IT and TC genotypes were heavier for the body weight at birth and at twenty weeks of age than those containing genotype. Cc. However, the allele T was significantly associated with higher eye muscle area(P < 0.05). As a result of this study, we suggested that the allele T in exon 3 of MSTN gene comes a significant effect for increasing the eye muscle area without decreasing backfat thickness. This polymorphism did not change the amino acid but Taq I -RFLP matched to SSCP band patterns in exon 3 of MSTN gene, which will be an useful molecular marker for breeding of Landrace pigs.

Development of a diagnostic method for human enteric Adenovirus-41 with rapid, specific and high sensitivity using the loop-mediated isothermal amplification assay

  • Lee, Jin-Young;Rho, Jae Young
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.673-681
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    • 2020
  • Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.

Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity (pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kang-Sun;Chung, Wee-Sup;Kim, Soo-Gwan;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.211-217
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    • 2006
  • Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

cDNA Cloning, Tissue Expression and Association of Porcine Pleiomorphic Adenoma Gene-like 1 (PLAGL1) Gene with Carcass Traits

  • Zhang, F.W.;Cheng, H.C.;Deng, C.Y.;Xiong, Y.Z.;Li, F.E.;Lei, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1257-1261
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    • 2006
  • Pleiomorphic adenoma gene-like1 (PLAGL1) encodes a zinc-finger (ZF) protein with seven ZFs of the C2H2-type which is a regulator of apoptosis and cell cycle arrest, and also regulates the secretion of insulin. In both human and mouse, PLAGL1 is a candidate gene for tumor suppressor and transient neonatal diabetes mellitus (TNDM). In this study, a 2,238 bp fragment covering the complete coding region was obtained and deposited to GenBank (accession number: DQ288899). The reverse transcriptase-polymerase chain reaction (RT-PCR) indicated that PLAGL1 was expressed almost equally in heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, uterus and ovary. Comparing the sequences of Large White and Meishan pigs, a C-T transition in exon 6 was found. The polymorphism could be detected by TaqI and was genotyped in five purebreds (Large White, Landrace, Meishan, Tongcheng and Bamei). Association analysis was performed between the polymorphism and carcass traits in 276 pigs of a "Large White${\times}$Meishan" F2 resource population. As a consequence, significant associations of the genotypes with shoulder backfat thickness (SFT) and internal fat rate (IFR) were observed. Pigs with TT genotype had low SFT and high IFR compared with TC or CC genotypes.

Effects on osteoporosis of Senescence Accelerated Mice (SAM) P6 by Injection of Placenta extracts ($Beuronel^{(R)}$) (SAM p6 mouse에서 자하거 주사제의 골다공증 개선에 대한 연구)

  • Kim, Mi-Jung;Yum, Do-Young;Koo, Bon-Taq;Ma, Jin-Yeul;Shin, Hyun-Kyu;Lee, Mi-Young;Shin, Dong-Won;Park, Jong-Cheol;Yu, Young-Beob
    • The Korea Journal of Herbology
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    • v.22 no.3
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    • pp.77-83
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    • 2007
  • Objectives : The purpose of this study was to examine the effects of the Hominis placenta extracts on improvement of senile osteoporosis (Type 2) in SAM P6. In traditional Oriental medicine, Placenta (Jahageo) is indicated for insufficiency of blood and tidal fever due to steaming bone disorder. Methods : At 10 weeks after birth, Placenta ex. was given 100 mL, 200 ml/kg/day, i.p. 3 times a week for 4 weeks in SAM P6 mice. And then we measured complete blood cells, blood urea nitrogen, creatinine, inorganic phosphate and total iron. In addition, we tested bone mineral density (BMD) using the soft X-ray. Results : As a result, in SAM P6, injection of Placenta ex. (200 mL) increased in RBC, HB and PLT, in comparison with control group. It was also found that the inorganic phosphate levels increased significantly in the injection groups of the Placenta ex. from that of the control group, but blood urea nitrogen was no significant. Moreover, Placenta ex. were showed a trend of increase in bone mineral density (BMD) of the vertebrae lumbales and tibia in P6 mice. Conclusions : These findings suggest that Placenta ex. are effective in preventing bone loss in SAMP6.

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Allelic Association of the Dopamine $D_2$ Receptor in Korean Alcoholics (한국인 알코올 중독 환자에서 도파민 $D_2$ 수용체의 대립유전자 연합)

  • Lee, Kang-Joon;Lee, Min-Soo;Kwak, Dong-Il
    • Korean Journal of Biological Psychiatry
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    • v.4 no.1
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    • pp.43-47
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    • 1997
  • The author attempted to examine the allelic association between the A1 allele of Dopamine $D_2$ receptor and alcoholism in Koreans. The allelic distribution of Taq I polymorphism of the $D_2$ dopamine receptor gene with alcoholism was examined in 67 Korean alcoholics and compared with 100 Korean controls. In alcoholics, the numbers of alcoholics with A1A1, A1A2 and A2A2 were 11(16.4%), 30(44.8%) and 26(38.8%) respectively and in controls with A1A1, A1A2 and A2A2 were 17(17.0%), 42(42.0%) and 41(41.0%), respectively. The prevalence of the A1 allele in alcoholics was 61.2% and 59.0% in controls. And the frequency of the A1 allele in alcoholics and controls were 0.39 and 0.38, respectively. There was not significant difference in the frequency of the A1 allele between alcoholics and controls. This data suggest that the A1 allele is not associated with alcoholism in Koreans. The author conclude that our data do not support an allelic association between the A1 allele at Dopamine $D_2$ receptor and alcoholism. Further systemized studies will be necessary to determine whether the role of allele of Dopamine $D_2$receptor is major effect gene or modifying effect gene in the pathogenesis of alcoholism.

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Simultaneous Molecular Detection of Cryptosporidium and Cyclospora from Raw Vegetables in Korea

  • Sim, Seobo;Won, Jua;Kim, Jae-Whan;Kim, Kyungjin;Park, Woo-Yoon;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.137-142
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    • 2017
  • Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and food-borne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, Cryptosporidium and Cyclospora were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, Cryptosporidium was isolated from all 6 kinds of vegetables, whereas Cyclospora was detected in 4 kinds of vegetables (except perilla leaves and chives). Cryptosporidium (17.8%) and Cyclospora (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. Cryptosporidium was detected all year long; however, Cyclospora was detected only from October to January. In 2 samples (sprout and blueberry), both Cryptosporidium and Cyclospora were detected. Further investigations using TaqI restriction enzyme fragmentation and nested PCR confirmed Cryptosporidium parvum and Cyclospora cayetanensis, respectively. In conclusion, we detected C. cayetanensis in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.

Molecular Characterization and Chromosomal Mapping of the Porcine AMP-activated Protein Kinase ${\alpha}2$ (PRKAA2) Gene

  • Lee, Hae-Young;Choi, Bong-Hwan;Lee, Jung-Sim;Jang, Gul-Won;Lee, Kyung-Tai;Chung, Ho-Young;Jeon, Jin-Tea;Cho, Byung-Wook;Lee, Jun-Heon;Kim, Tae-Hun
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.615-621
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    • 2007
  • AMP-activated protein kinase alpha 2 (PRKAA2) plays a key role in regulation of fatty acid and cholesterol metabolism. This study investigated the porcine PRKAA2 gene as a positional candidate for intramuscular fat and backfat thickness traits in pig chromosome 6. A partial fragment of the porcine PRKAA2 gene, amplified by PCR, contained a putative intron 3 including a part of exon 3 and 4, comparable with that of human PRKAA2 gene. Within the fragment, several single nucleotide polymorphisms were identified using multiple sequence alignments. Of these, TaqI restriction enzyme polymorphism was used for genotyping various pig breeds including Korean reference family. Using linkage and physical mapping, the porcine PRKAA2 gene was mapped in the region between microsatellite markers SW1881 and SW1680 on chromosome 6. Allele frequencies were quite different among pig breeds. The full length cDNA of the porcine PRKAA2 (2,145 bp) obtained by RACE containing 1,656 bp open reading frame of deduced 552 amino acids, had sequence identities with PRKAA2 of human (98.2%), rat (97.8%), and mouse (97.5%). These results suggested that the porcine PRKAA2 is a positional candidate gene for fat deposition trait at near telomeric region of the long arm of SSC 6.

Characterization of porcine cytokine inducible SH2-containing protein gene and its association with piglet diarrhea traits

  • Niu, Buyue;Guo, Dongchun;Liu, Zhiran;Han, Xiaofei;Wang, Xibiao
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.12
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    • pp.1689-1695
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    • 2017
  • Objective: The cytokine inducible SH2-containing protein (CISH), which might play a role in porcine intestine immune responses, was one of the promising candidate genes for piglet anti-disease traits. An experiment was conducted to characterize the porcine CISH (pCISH) gene and to evaluate its genetic effects on pig anti-disease breeding. Methods: Both reverse transcription polymerase chain reaction (RT-PCR) and PCR were performed to obtain the sequence of pCISH gene. A pEGFP-C1-CISH vector was constructed and transfected into PK-15 cells to analysis the distribution of pCISH. The sequences of individuals were compared with each other to find the polymorphisms in pCISH gene. The association analysis was performed in Min pigs and Landrace pigs to evaluate the genetic effects on piglet diarrhea traits. Results: In the present research, the coding sequence and genomic sequence of pCISH gene was obtained. Porcine CISH was mainly localized in cytoplasm. TaqI and HaeIII PCR restriction fragment length polymorphism (RFLP) assays were established to detect single nucleotide polymorphisms (SNPs); A-1575G in promoter region and A2497C in Intron1, respectively. Association studies indicated that SNP A-1575G was significantly associated with diarrhea index of Min piglets (p<0.05) and SNP A2497C was significantly associated with the diarrhea trait of both Min pig and Landrace piglets (p<0.05). Conclusion: This study suggested that the pCISH gene might be a novel candidate gene for pig anti-disease traits, and further studies are needed to confirm the results of this preliminary research.

Association of Melanocortin 4 Receptor (MC4R) Genotypes with Fatty Acid Compositions in an Intercross Population between Duroc and Jeju Native Pigs (제주재래돼지와 듀록 참조축군에서 Melanocortin 4 Receptor (MC4R) 유전자형과 지방산 조성간의 관련성 분석)

  • Kang, Yong-Jun;Kim, Sang-Geum;Kim, Su-Yeon;Shin, Moon-Cheol;Woo, Jae-Hoon;Kim, Nam-Young;Shin, Sang-Min;Choi, Jae-Young;Yoo, Ji-Hyun;Park, Nam-Geon;Yang, Byoung-Chul;Cho, In-Cheol
    • Journal of Life Science
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    • v.30 no.1
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    • pp.58-63
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    • 2020
  • This study was conducted to examine association between melanocortin-4-receptor (MC4R) genotypes and fatty acid (FA) composition in an F2 intercross between Duroc and Jeju (South Korea) Native pigs (JNP). Fourteen FA composition traits were measured in more than 290 F2 progeny population produced between Duroc and JNP. All experimental pigs were successfully genotyped for the MC4R c.1426A>G (p.Asp298Asn) single nucleotide polymorphism (SNP) by using Taq I PCR-RFLP methods. We detected three MC4R genotypes, AA, AG, and GG with 0.299, 0.542, and 0.159 genotype frequencies, respectively. The MC4R AA genotype animals showed higher levels in palmitic acid (C16:0, p<0.05), stearic acid (C18:0, p<0.01), eicosenoic acid (C20:1n9, p<0.05), saturated fatty acid (SFA, p<0.01) than GG homozygotes, respectively. Whereas MC4R GG genotype showed higher values in linoleic acid (C18:2n6, p<0.001), linolenic acid (C18:3n3, p<0.001), linolenic acid (C18:3n6, p<0.001), arachidonic acid (C20:4n6, p<0.001) and unsaturated fatty acid (USFA, p<0.01) than AA pigs, respectively. The MC4R GG genotype was associated with increasing USFA and decreasing SFA in the F2 progeny population produced between Duroc and JNP. Our finding suggests that the MC4R polymorphisms can be used as a valuable genetic marker for Duroc and JNP breeding programs to improve meat quality and to control FA compositions.