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PCR-RFLP for the Identification of Mammalian Livestock Animal Species
Han, Sang-Hyun,Park, Seon-Mi,Oh, Hong-Shik,Kang, Geunho,Park, Beom-Young,Ko, Moon-Suck,Cho, Sang-Rae,Kang, Yong-Jun,Kim, Sang-Geum,Cho, In-Cheol The Korean Society of Embryo Transfer 2013 한국동물생명공학회지 Vol.28 No.4
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.
( Sehun Kim ),( Donghoon Han ),( Jae Hyuk Choi ),( Eun-joo Park ),( Dong Geum Shin ),( Min-kyung Kang ),( Seonghoon Choi ),( Namho Lee ),( Jung Rae Cho ) 대한내과학회 2021 The Korean Journal of Internal Medicine Vol.36 No.0
Background/Aims: Prolonged dual antiplatelet therapy (DAPT) with aspirin and clopidogrel beyond 1 year has been shown to reduce ischemic events at the expense of increased bleeding. However, limited data are available on the clinical significance of platelet reactivity (PR) at 1 year. Methods: We retrospectively identified 331 patients who underwent percutaneous coronary intervention (PCI) and assessed the on-clopidogrel PR using VerifyNow P2Y12 assay at 1 year in a single center. Two hundred eleven patients were on DAPT for > 1 year. The relationship between high on-treatment platelet reactivity (HPR) at 1 year and clinical outcomes beyond 1 year, as well as the longitudinal change in PR was analyzed. Results: At 1 year, 135 (64%) patients showed HPR and 76 (36%) did not. There was a significant increase in ischemic endpoint events, including cardiovascular death, non-fatal myocardial infarction, and stroke/transient ischemic attack in patients with compared to without HPR at 1 year (hazard ratio [HR], 2.68; 95% confidence interval [CI], 1.06 to 6.77; p = 0.036). However, the incidence of any Bleeding Academic Research Consortium bleeding was significantly lower in the HPR group (HR, 0.11; 95% CI, 0.02 to 0.65; p = 0.015). In the longitudinal analysis, PR significantly decreased from post-load to 1 year after index PCI in the non-HPR group. Conversely, the HPR group showed high PR from baseline through 1 year. Conclusions: HPR at 1 year may be a useful surrogate for predicting ischemic and bleeding events in patients on prolonged DAPT. Patients with and without HPR at 1 year showed different patterns of longitudinal change in PR.
PCR-RFLP for the Identification of Mammalian Livestock Animal Species
Sang-Hyun Han,Seon-mi Park,Hong-Shik Oh,Geunho Kang,Beom-Young Park,Moon-Suck Ko,Sang-Rae Cho,Yong-Jun Kang,Sang-Geum Kim,In-Cheol Cho 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.4
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.
제주흑우, 한우 및 수입 소 품종에서 새로운 indel 마커의 다형성과 대립인자 분포
한상현(Sang-Hyun Han),김재환(Jae-Hwan Kim),조인철(In-Cheol Cho),조상래(Sang-Rae Cho),조원모(Won-Mo Cho),김상금(Sang-Geum Kim),김유경(Yoo-Kyung Kim),강용준(Yong-Jun Kang),박용상(Yong-Sang Park),김영훈(Young-Hoon Kim),박세필(Se-Phil Pa 한국생명과학회 2012 생명과학회지 Vol.22 No.12
본 연구는 소 유전자 database들에 대한 사전 비교연구에서 발견된 삽입/결실(indel) marker들의 다형성과 각각의 유전자형의 분포를 확인하고자 수행하였다. 먼저, 소의 유전체 서열과 발현서열표식(EST) database 간의 생물정보학적 비교를 통해 전체 51 종의 indel marker들을 검출하였다. 이 중에서 42 종을 평가하여 최종적으로 9 종의 정보력이 있는 marker들을 집단분석을 위해 선발하였다. 각각의 marker들에 대한 염기서열을 재분석하였으며, marker의 다형성을 한국 재래소 품종인 한우와 제주흑우(JBC), Holstein, Angus, Charolais, Hereford 등 6 품종에서 조사하였다. 본 연구에서 이용한 소 6 품종은 8 종의 marker들에 대해 다형성을 나타내었으나, Indel_15의 경우 Holstein과 Charolais에서 다형성이 발견되지 않았다. JBC 집단에 대한 분석에서는 관찰된 이형 접합자 빈도는 HW_G1 (0.600)에서 가장 높고, Indel_29 (0.274)에서 가장 낮았다. Marker에 대한 다형정보량의 수준은 HW_G4 (0.373)에서 가장 높고, Indel_6 (0.305)에서 가장 낮은 수준을 보였다. 본 연구에서 조사한 새로운 indel marker들은 특히 제주흑우 집단의 생산성 향상을 위한 분자육종 체계의 개발뿐만 아니라 친자확인이나 생산이력추적을 위한 유전정보를 제공하는데 유용할 것으로 기대된다. The aim of this study was to screen the polymorphisms and distribution of each genotype of insertion/deletion (indel) markers which were found in a preliminary comparative study of bovine genomic sequence databases. Comparative bioinformatic analyses were first performed between the nucleotide sequences of Bovine Genome Project and those of xpressed sequence tag (EST) database, and a total of fifty-one species of indel markers were screened. Of these, forty-two indel markers were evaluated, and nine informative indel markers were ultimately selected for population analysis. Nucleotide sequences of each marker were re-sequenced and their polymorphic patterns were typed in six cattle breeds: Holstein, Angus, Charolais, Hereford, and two Korean native cattle breeds (Hanwoo and Jeju Black cattle). Cattle breeds tested in this study showed polymorphic patterns in eight indel markers but not in the Indel-15 marker in Charolais and Holstein. The results of analysis for Jeju Black cattle (JBC) population indicated an observed heterozygosity (Ho) that was highest in HW_G1 (0.600) and the lowest in Indel_29 (0.274). The PIC value was the highest in HW_G4 (0.373) and lowest in Indel_6 (0.305). These polymorphic indel markers will be useful in supplying genetic information for parentage tests and traceability and to develop a molecular breeding system for improvement of animal production in cattle breeds as well as in the JBC population.