• Biotechnology and Bioprocess Engineering:BBE
• /
• 제5권2호
• /
• pp.71-78
• /
• 2000

The recent progress od DNA technologies including DNA fingerprinting (DFP) and random amplified DNA polymorphism (RAPD) analysis make it possible to identify the specific genetic trits of animals and to analyze the genetic diversity and relatedness between or withinspecies or populations. Using those techniquse, some efforts to identify and develop the specific DNA markers based on DNA polymorphism, which are related with economic traits for Korean native animals, Hanwoo(Korean native cattle),Korean native pig and Korean native chicken, have been made in Korea for recent a few years. The developed specific DNA markers successfully characterize the Korean native animals as the unique Korean genetic sources, distinctively from other imported breeds. Some of these DNA markers have been related to some important economic traits for domestic animals, for example, growth rate and marbling for Honwoo, growth rate and back fat thinkness fornative pig, and growth rate, agg weight and agg productivity for native chicken. This means that those markers can be used in important marker-assised selection (MAS) of Korean native domestic animals and further contribute to genetically improve and breed them.

• 통합자연과학논문집
• /
• 제1권3호
• /
• pp.230-233
• /
• 2008

A simple and rapid method is described for extending the 5'- or 3'-end genomic sequence of a known partial sequence by only a single round of PCR. This method involves digesting and ligating genomic and plasmid DNAs, and amplifying the 5'-upstream or 3'-end downstream sequence of the known DNA sequence, using two primers, one gene specific and the other plasmid specific. A single round of PCR amplification is sufficient to produce gene-specific bands detectable in gels. By using this approach, 5'-end genomic sequence of the D-amoeba sams gene was extended.

• 미생물학회지
• /
• 제39권2호
• /
• pp.114-117
• /
• 2003

유전자의 기능을 분석하는 과정에서 특정한 염기 서열의 존재여부를 확인하는 분석법은 필수적이다. 현재 사용되고 있는 Southern및 Northern blotting방법은 시간이 오래 걸리며, 온도 등과 같은 외부 조건을 엄격하게 조절하여야 한다. 본 연구에서는 측방유동방식을 이용한 크로마토그라피법을 응용하여 새로운 간편용 DNA분석법을 개발하였다. 이 측방유동형 DNA 분석 스트립은 시료가 적용되는 샘플패드, 이동하여 분리되고 교잡반응이 일어나는 전개용 막, 그리고 시료가 계속하여 이동하기 위한 흡수패드로 구성되어 있다. 모델 시스템으로 HIV와 HCV에 대한 포획 및 표적 DNA를 합성하고 스트립을 제조하였다. 시료를 샘플패드에 적하한 후 교잡반응체의 생성여부와 상대적인 양은 GSI형광 스캐너로 분석하였다. 교잡반응이 매우 빠르게 진행되고 세척과정이 없음에도 불구하고 비특이적인 교차 반응이 거의 관찰되지 않았다. 기존의 DNA 교잡방법과 비교하여 볼 때 이 새로운 방법으로 DNA/DNA 교잡 실험을 보다 더 쉽고, 간편하고, 그리고 빠르게 할 수가 있을 것으로 예상된다.

• Restorative Dentistry and Endodontics
• /
• 제24권1호
• /
• pp.13-25
• /
• 1999

P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

• International Journal of Oral Biology
• /
• 제46권4호
• /
• pp.155-159
• /
• 2021

This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249^T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249^T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

• Journal of Microbiology and Biotechnology
• /
• 제16권5호
• /
• pp.740-747
• /
• 2006

Amplification by universal consensus sequences in pathogenic bacterial DNA would allow rapid identification of pathogenic bacteria, and amplification of genus-specific and species-specific sequences of pathogenic bacterial DNA might be used for genotyping at the genus and species levels. For design of probes for molecular diagnostics, several tools are available as stand-alone programs or as Web application. However, since most programs can design only a few probe sets at one time, they are not suitable for large-scale and automatic probes design. Therefore, for high-throughput design of specific probes in diagnostic array development, an automated design tool is necessary. Thus, we developed a Web-based automatic system for design of genus-specific and species-specific probes for pathogen detection. The system is available at http://www.miprobe.com.

• Clinical and Experimental Reproductive Medicine
• /
• 제46권4호
• /
• pp.147-151
• /
• 2019

Objective: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST). Methods: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern ("a"-"g") were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed. Results: The HOST examinations showed that > 93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type "d", 98.67% in type "g", and 98.17% in type "f" spermatozoa. Conclusion: We found that the type "d" spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.

• 분석과학
• /
• 제12권1호
• /
• pp.80-83
• /
• 1999

사람에게서 유래된 DNA시료의 X,Y 특이의 alphoid gene을 PCR법으로 분석하면 성별을 확인할 수 가 있다. PCR법으로 alphoid gene을 분석한바 매우 예민도가 높아 genomic DNA 약 60pg까지 성별을 분석할 수 있었다. 그리고 성별이 혼합되어 있는 DNA에서 female DNA의 1/10비 까지는 male DNA를 분석할 수 있었다. 따라서 이 결과는 혼합된 DNA에서 X,Y 특이의 alphoid gene을 분석하는데 기준으로 활용할 수가 있다.

• 대한기계학회논문집A
• /
• 제28권8호
• /
• pp.1135-1139
• /
• 2004

This paper presents a microextractor for the separation of DNA molecules by their sizes. The DNA microextractor immobilizes the DNA molecules of specific size in the micropillar array by adjusting the period of the crossed electric field, thus providing a starting-point independent target DNA extraction method without separation process monitoring. The DNA microextractor has been fabricated by a three-mask micromachining process. The velocity of three different DNA molecules has been measured at the electric field of E=5V/0.8cm in the fabricated DNA microextractor, resulting in the reorientation times of $4.80{\pm}0.44sec,\;7.12{\pm}0.75sec$, and $9.88{\pm}0.30sec$ for ${\lambda}$ DNA, micrococcus DNA, and T4 DNA, respectively. T4 DNA is trapped in the micropillar array when the crossed electric field of 5V/0.8cm is applied alternately at a 10 second time interval. The present DNA microextractor filters the DNA in a specific size range by adjusting the magnitude and/or the period of the crossed electric field applied in the micropillar array.

• Journal of Animal Science and Technology
• /
• 제50권1호
• /
• pp.9-18
• /
• 2008

본 연구에서는 조류에서 종 특이적인 DNA 염기서열변이를 검증하기 위하여 닭, 꿩, 칠면조, 메추리의 immunoglobulin light chain constant domain 유전자를 클로닝하여 DNA염기서열을 분석하였다. 종간에 구별이 가능한 DNA 염기서열변이가 위의 유전자에서 관찰되었다. PCR을 이용하여 종을 구별하기 위하여 종 사이에 특이적인 DNA 염기서열 부위에 한 쌍의 종 특이적인 프라이머를 제작하였다. 또한 비교실험을 위하여 이미 알려진 cytochrome b와 tapasin 유전자에서도 두 쌍의 종 특이적 프라이머를 제작하였다. PCR결과 세 쌍의 프라이머 모두 종 특이적으로 DNA를 증폭하였다. Immunoglobulin 유전자의 염기서열 변이를 이용한 종 특이적인 PCR 방법은 조류의 유전자원 보존을 위한 이종간 카이메라 연구에 유용하게 이용될 수 있을 것이다.