• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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• pp.177-180
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• 2004

본 연구는 PCR-RFLP 및 PCR-SSCP 기법을 이용하여 PSE 돈육을 생산하는 PSS 돼지 유전자 진단 기술을 개발하고 이를 이용한 국내 종돈 및 교잡 비육돈의 PSS 유전자 출현 빈도를 파악하고자 수행하였다. 돼지 PSS의 원인이 되는 ryanodine receptor 유전자의 단일염기 돌연변이 $C{\rightarrow}T$ ; $Arg\;{\rightarrow}\;Cys$)를 포함하는 134 bp 영역을 PCR로 증폭한 후 RFLP 및 SSCP 기법으로 분석한 결과 동형접합체의 정상(N/N), 이형접합체의 잠재성 개체 (N/n) 그리고 돌연변이 유전자를 동형접합체 상태로 갖는 PSS 감수성 개체(n/n)에 각각 특이적인 유전자형이 검출되었다. 특히, PCR-SSCP기법을 이용한 RYR1 유전자 돌연변이 검출 방법은 보다 신속 간편하면서도 상대적으로 분석비용이 저렴한 정확성이 높은 PSS 돼지 진단기술로서 대규모 돼지집단검색이나 RFLP 방법으로 판정이 불확실한 시료의 재검에 효율적으로 이용할 수 있을 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4319-4323
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• 2014

Background: Phospholipase C epsilon 1 (PLCE1) encodes a member of the phospholipase family of proteins that play crucial roles in carcinogenesis and progression of several cancers including esophageal cancer (EC). In two large scale genome-wide association studies (GWAS) single nucleotide polymorphisms (SNP, rs2274223A>G, rs3765524C>T) in PLCE1 were identified as novel susceptibility loci of esophageal cancer (EC) in China. The aim of the present study was to investigate this finding in Kashmir Valley, a high risk area. Materials and Methods: We determined genotypes of three potentially functional SNPs (rs2274223A>G, rs3765524C>T and rs7922612C>T) of PLCE1 in 135 EC patients, and 195 age and gender matched controls in Kashmiri valley by PCR RFLP method. Risk for developing EC was estimated by binary logistic regression using SPSS. Results: The selected PLCE1 polymorphisms did not show independent association with EC. However, the $G_{2274223}T_{3765524}T_{7922612}$ haplotype was significantly associated with increased risk of EC (OR=2.92; 95% CI=1.30-6.54; p=0.009). Smoking and salted tea proved to be independent risk factors for EC. Conclusions: Genetic variations in PLCE1 modulate risk of EC in the high risk Kashmiri population.

• Parasites, Hosts and Diseases
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• 제53권4호
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• pp.493-496
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• 2015

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

• 한국미생물·생명공학회지
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• 제36권4호
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• pp.247-259
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• 2008

미생물 구조 및 집단 구성원의 변화를 배양에 의존하지 않고, 빠르게 평가할 수 있는 방법의 하나인 terminal-restriction fragment length polymorphism (I-RFLP) 분석을 김치 유산균 연구에 적용하여, $15^{\circ}C$ 및 $4^{\circ}C$ 발효 김치에 관여하는 유산균의 다양성과 역동성을 검토하였다. 두 발효온도에서 공통적으로 Leuconostoc mesenteroides, Lc, inhae, Lc. kimchii, Weissella koreensis, W. cibaria, Lactobacillus sakei, Lb. curvatus, Lb. plantarum, Lb. paraplantarum, Lb. pentosus, 및 Lb. brevis의 존재가 예상되었고, Lc. citreum과 Enterococcus faecalis는 각각 $15^{\circ}C$와 $4^{\circ}C$에서 검출되었다. W. koreensis는 중기발효에 우점을 점하였고, Lactobacillus 속은 발효 후기의 우점종으로 나타났다. Lb. sakei, Lb. curvatus는 발효온도에 관계 없이 우점종을 형성하였지만, Lb. plantarum, Lb. paraplantarum, Lb, pentosus와 Lb. brevis의 생육은 저온에서 잘 일어나지 않았다. 몇 종의 Leuconostoc 속 유산균은 발효 후기까지 생장이 유지되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1095-1099
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• 2006

Leptin and IGFBP-3 are two proteins that play an important role in growth and metabolism of the animals. They are also involved in the immune function of animals and, thus, are candidate genes for the study of association with immune functions. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of these two genes was done to screen 64 crossbred (Holstein Friesian${\times}$Hariana) female calves of one year of age. From each RFLPs (fragments) three genotypes were observed. In all the RFLPs the mutant homozygotes were very less in numbers and, hence, were excluded from the least squares analysis. The serum IgG level was estimated using SRID assay. The mean level of serum IgG was $28.83{\pm}2.73mg/ml$. The effect of these identified genotypes on serum IgG level of calves at one year of age was analysed using least squares analysis. The HaeIII RFLP-AB genotype had significantly (p<0.05) higher serum IgG level ($31.86{\pm}3.05$) than the HaeIII RFLP-AA ($25.62{\pm}2.96$) genotype. There was no significant effect of leptin genotypes on the IgG level. The present results indicated a role of the IGFBP-3 gene on serum IgG level of cattle calves.

• Interdisciplinary Bio Central
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• 제4권3호
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• pp.8.1-8.7
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• 2012

The motor neuron degeneration 2 (mnd2) mice carry a point mutation of A to T nucleotide transversion at the serine 276 residue of high temperature requirement A2 (HtrA2), resulting in losses of an AluI restriction enzyme site (5'AGCT3') and the HtrA2 serine protease activity. Moreover, dysfunctions of HtrA2 are known to be intimately associated with the pathogenesis of neurodegenerative diseases, including Parkinson's disease. Thus, this mnd2 mouse is an invaluable model for understanding the physiological role of HtrA2 and its pathological role in neurodegenerative diseases. Nevertheless, many molecular and cellular biologists in this field have limited experience in working with mutant mouse models due to the necessity of acquired years of the special techniques and knowledges. Herein, using the mnd2 mouse model as an example, we describe easy-to-use standard protocols for web-based analyses of target genes, such as HtrA2, and a novel approach for simple and accurate PCR-AluI-RFLP-based genotype analysis of mnd2 mice. In addition, band resolution of AluI-RFLP fragments was improved in 12% polyacrylamide gel running in 1X Tris-Glycine SDS buffer. Our study indicates that this PCR-AluI-RFLP genotype analysis method can be easily applied by the molecular and cellular biologist to conduct biomedical science studies using the other mutant mouse models.

• 한국가금학회지
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• 제41권3호
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• pp.191-196
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• 2014

가금 사육 프로그램에서 경제적으로 중요한 형질에 잠재적인 후보 유전자의 식별 및 활용은 점점 더 중요해지고 있다. Ovocalyxin-32(OCX-32) 유전자는 닭의 9번 염색체에 위치하며, 난각을 형성하는데 중요한 역할을 한다. 본 연구의 목적은 한국 재래닭의 OCX-32 유전자 내 SNP의 유전자형 결정과 산란 형질과의 연관성을 분석하기 위해 수행하였다. PCR-RFLP 방법을 통해 한국 오골계 46수, 백색 46수, 회색 43수, 흑색 46수를 포함한 총 181수의 한국 재래닭 암컷 4종의 SNP을 분석하였다. 산란 형질은 시산일령, 시산난중, 산란율, 난중의 4가지 항목을 포함하여 측정하였다. OCX-32 유전자 내 c.494A>C SNP은 오골계의 산란율과 유의적인 차이를 나타냈으며(p<0.001), 백색 재래닭에서는 난중과 유의적인 상관관계가 있었다(p<0.05). c.267T>G SNP은 오골계의 난중과 유의적인 연관성이 나타났다(p<0.05). 하지만 회색과 흑색 재래닭에서는 유의적인 상관관계가 나타나지 않았다. 본 연구 결과, 한국 재래닭의 사육 프로그램에서 산란형질을 선발하는 마커로 사용되기까지 추후 더 많은 개체군을 통한 연구가 요구되나, OCX-32 유전자 내 c.494A>C와 c.267T>G의 단일염기변이가 한국 재래닭 중 오골계와 백색 재래닭에서 산란 특성에 따른 DNA 선발 마커로서 활용할 수 있을 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제55권3호
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• pp.239-246
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• 2017

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of $Ilh{\acute{e}}us$ and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.196.4-196
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• 2005

• Journal of Microbiology and Biotechnology
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• 제13권1호
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• pp.15-21
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• 2003

Two microbial communities of activated sludge in the same municipal wastewater, but treated with different systems, were studied and compared using molecular microbiological approaches. The bacterial 16S rDNA sequences from 124 clones were analyzed, however, the majority of them were not closely related to any known species, and found to belong to 8 different phylogenetic groups and 3 different unidentified groups. The relative frequencies of each group were similar between the two microbial communities. Fingerprinting using terminal restriction fragment length polymorphism (T-RFLP) showed that the putative Nitrospira-related populations were more diverse and quantitatively higher in the KNR process system than in the other system using a conventional activated sludge process. The relationship between the bacterial community composition and the higher removal efficiency of nitrogen and phosphorus in the KNR process is discussed.