• Title/Summary/Keyword: primer

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Genotyping of HLA-A by Polymerase Chain Reaction-Sequence Specific Primer (Polymerase Chain Reaction-Sequence Specific Primer를 이용한 HLA-A 유전자의 DNA 다형성 조사)

  • Jang, Soon-Mo
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.2
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    • pp.94-97
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    • 2008
  • The human leukocyte antigen (HLA) is the name of the major histocompatibility complex (MCH) in humans. The superlocus contains a large number of genes related to immune system function in humans. This group of genes resides on chromosome 6. and encode cell surface antigen-presenting proteins and many other genes. HLA class I antigen (A, B & C) present peptides from inside the cell. These peptides are produced from digested proteins that are broken down in the lysozymes. Most expressed HLA loci exhibit a remarkable degree of allelic polymorphism, which derives from sequence differences predominantly localized to discrete hypervariable regions of the amino terminal domain of the molecule. In this sutdy, the HLA-A genotypes were determined in twenty students unrelated koreans using the PCR-SSP (Polymerase Chain Reaction-Sequence Specific Primer) technique. Several specific primer pairs in assigning the HLA-A gene were used (A*0201, A*33, A*2401). The results of PCR-SSP, the HLA-A*0201 primer was detected eleven (55%), the HLA-A*33 were detected seven (35%) and the HLA-A*2401 were detected seven (35%). This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-A genotypes.

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Multiplex PCR Assay for Simultaneous Detection of Korean Quarantine Phytoplasmas

  • Kim, Young-Hwan;Win, Nang Kyu;Back, Chang-Gi;Yea, Mi-Chi;Yim, Kyu-Ock;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.367-371
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    • 2011
  • Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

ITS Primers with Enhanced Specificity to Detect the Ectomycorrhizal Fungi in the Roots of Wood Plants

  • Kim, Dong-Hun;Chung, Hung-Chae;Ohga, Shoji;Lee, Sang-Sun
    • Mycobiology
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    • v.31 no.1
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    • pp.23-31
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    • 2003
  • With universal primer ITS1-F, the specific DHJ2 primer was developed to detect the Ectomycorrhizal(ECM) root tips in soil and to identify the species of ECM fungi, as based on DNA sequences of rDNA stored in GeneBank of NCBI. This primer was designed with the common sites of rDNA of Amanita and Boletus, and was also designed with several DNA programs provided by NCBI. The DNA fragments synthesized by PCR were calculated to be 1,000 to 1,200 bps of DNA located to 18s to 28s rDNA to contain two variable sites of ITS, indicating much diversities for specific species or ecotypes of ECM fungi. The primer DHJ2 reacted with the genomic DNA's extracted from the tissues of basidiocarp at the rate of 73 of 80 fungi collected produced single bands with a 1,100 bps length. The DNA fragment synthesized with the genomic DNA that extracted from eight ECM tips of Pinus densiflora was confirmed and analysized to the rDNAs of ECM in full sequences, and informed to be a ECM fungal species in the forest.

Genotyping of HLA-B by Polymerase Chain Reaction-Sequence Specific Primer (Polymerase Chain Reaction-Sequence Specific Primer를 이용한 HLA-B 유전자의 DNA 다형성 조사)

  • Jang, Soon-Mo
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.147-150
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    • 2007
  • Most expressed HLA (human leukocyte antigen) loci exhibit a remarkable degree of allelic polymorphism, which derives from sequence differences predominantly localized to discrete hypervariable regions of the amino terminal domain of the molecule. In this study, the HLA-B genotypes were determined in twenty students unrelated koreans using the PCR-SSP (polymerase chain reaction-sequence specific primer) technique. Several specific primer pairs in assigning the HLA-B gene were used ($B^{\ast}4001/4007$, $B^{\ast}4901/5001/4501$, $B^{\ast}3701$, $B^{\ast}5801$). The results of PCR-SSP, the HLA-B3701 primer was detected one (5%), the $HLA-B^{\ast}5801$ were detected four (20%), the $HLA-B^{\ast}4001/4007$ were detected nineteen (95%) and the $HLA-B^{\ast}4901/5001/4501$ were detected twenty. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-B genotypes. Moreover, these results genotype frequency of the HLA-B gene could be useful for database study before being applied to individual identification and transplantation immunity.

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

An Analysis of the Cause of Porosity Generation and Reduction Plan in Fillet Welding (필렛용접에서의 결함발생 원인 분석 및 저감 방안)

  • Choi, K.Y.;Kim, Y.P.;Kim, K.J.;Kim, D.S.;Bae, S.D.
    • Special Issue of the Society of Naval Architects of Korea
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    • 2006.09a
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    • pp.128-133
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    • 2006
  • Generally, porosity which was formed by pyrolysis of the primer is usually generated in the weld metal in respect of increase of the welding speed. in order to analyze the cause of porosity generation, this study was performed using FCAW(flux cored arc welding) process for three kinds of inorganic.zinc primer. in addition the evaluation by influence of welding method on porosity generation is conducted to compare between FCAW and MAG(metal active gas) welding with the same inorganic zinc primer. As the result of this investigation, not only primer of lower organic binder and zinc but also FCAW process than MAG in fillet welding have been verified the excellent resistance to the porosity generation for horizontal fillet welding.

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Identification of AFLP Marker Linked to a SCN Resistant Gene in Soybean

  • Ko, Mi-Suk;Kim, Myung-Sik;Han, Soung-Jin;Chung, Jong-Il;Kang, Jin-Ho
    • Plant Resources
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    • v.5 no.3
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    • pp.169-175
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    • 2002
  • The soybean cyst nematode (Heterodera glycines Inchinoe; SCN) is a devastating pest of soybean and is responsible for significant losses in yield. The use of resistant cultivars is the effective method to reduce or eliminate SCN damage. The objective of this research is to identify AFLP markers linked to the SCN resistant genes. Bulked genomic DNA was made from resistant and susceptible genotypes to SCN and a total of 19 primer combinations were used. About 31 fragments were detected per primer combination. The banding patterns were readily distinguished in resistant and susceptible bulked genotypes. Polymorphic fragments were detected between resistant and susceptible bulked genotypes in the primer combination of CGT/GGC, CAG/GTG and CTC/GAG. In primer combinations of CGT/GGC and CAG/GTG, bulked resistant genotype produced a polymorphic bands. However, in primer of CTC/GAG, bulked susceptible genotype produced a polymorphic fragments. Three AFLP markers identified as a polymorphic fragments between bulked genomic DNA were mapped in 85 F2 population. Among them, only two markers, CGT/GGC and CTC/GAG, was linked and was mapped. Broad application of AFLP marker would be possible for improving resistant cultivars to SCN.

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Comparison of RAPD Profiles and Phenotypical Characters of Streptococcal Strains (연쇄상구균의 표현형적 특성과 RAPD profiles 비교)

  • Song, Jin-Gyeong;Kim, Jong-Hun;Kim, Eun-Hui
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.51-59
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    • 2003
  • Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

Identification of Nocardia seriolae by polymerase chain reaction (PCR에 의한 Nocardia seriolae의 검출)

  • Park, Myoung-Ae;Cho, Mi-Young;Kim, Myoung-Sug;Kim, Jae-Hoon;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.1
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    • pp.85-90
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    • 2009
  • A method for the identification of Nocardia seriolae, the causative agent of nocardiosis in cultured fishes, using PCR was developed in the study. A PCR primer set specific to N. seriolae was designed based on 16S-23S rRNA sequence of various Nocardia species accessed in GenBank. Designed PCR primer set, Nseri-F (5'-GCA AAC TCT TCG AAC AGT CG-3') and Nseri-R (5'-GGA TAT CAG GAC TTA CCG GC-3'), amplifies the target regions of N. seriolae only, but not 4 other Nocardia species, N. asteroides, N. crassostreae, N. farcinica and N. salmonicida.

Design and Implementation of a PCR Primer Search System on Cloud Computing Environments (클라우드 컴퓨팅 환경에서 PCR Primer 검색 시스템 설계 및 개발)

  • Park, Junho;Lim, Jongtae;Kim, Dongjoo;Lee, Yunjeong;Ryu, Eunkyung;Ahn, Minje;Cha, Jaehong;Yu, Seok Jong;Yoo, Jaesoo
    • Proceedings of the Korea Contents Association Conference
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    • 2012.05a
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    • pp.269-270
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    • 2012
  • 유전자 증폭을 위한 정확한 PCR Primer의 디자인은 핵심적인 기반 기술이다. 기존 연구를 통해 각 유전자별 특이적인 PCR Primer를 디자인할 수 있는 도구가 제안되었으나, 유전체 정보를 활용한 대단위의 디자인작업을 수행하기에는 적합하지 않았다. 본 논문에서는 클라우드 컴퓨팅 환경에서 대규모의 유전체를 대상으로 특이적인 PCR Primer를 디자인하고 검색할 수 있는 시스템을 설계하고 구현한다. 제안하는 시스템은 Hadoop 플랫폼에서의 MapReduce 프레임워크를 기반으로 설계 및 구현하여 유전자 서열검색을 대규모로 수행할 수 있도록 하였다. 5만개의 질의를 이용한 성능 평가 결과, 제안하는 기법은 기존 BLAST를 이용한 검색방법에 비해 약 3배의 성능 향상을 보였다.

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