• Animal cells and systems
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• 제5권2호
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• pp.153-156
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• 2001

A key aspect of genomic research in the “post-genome era”is to associate sequence variations with heritable phenotypes. The most common variations in the human genome are single nucleotide polymorphisms (SNPs) that occur approximately once in every 500 to 1,000 bases. Although analyzing the phenotypic outcome of these SNPs is crucial to facilitate large-scale association studies of genetic diseases, detection of SNPs from an extended number of human DNA samples is often difficult, labor-intensive and time-consuming. Recent development in SNP detection methods using DNA microarrays and mass spectrophotometry has allowed automated high throughput analyses, but such equipments are not accessible to many scientists. In this study, we demonstrate that a simple PCR-based method using primers with a mismatched base at the 3'-end provides a fast and easy tool to identify known SNPs from human genomic DNA in a regular molecular biology laboratory. Results from this PCR amplification of specific alleles (PASA) analysis efficiently and accurately typed the Q576R polymorphism of human IL4 receptor from the genomic DNAs of 29 Koreans, including 9 samples whose genotype could not be discerned by the conventiona1 PCR-SSCP (single strand conformation polymorphism) method. Given the increasing attention to disease-associated polymorphisms in genomic research, this alternative technique will be very useful to identify SNPs in large-scale population studies.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2005년도 제36차 추계 학술발표대회
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• pp.123-126
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• 2005

본 연구는 고등동물의 불포화지방산 생합성의 핵심 효소로 알려져 있는 Stearoyl-CoA desaturase(SCD) 유전자의 특정한 단일염기다형(single nucleotide polymorphism; SNP)이 한우의 도체 및 육질형질에 미치는 영향을 분석하기 위해 SCD 유전자의 Intron 7번 영역의 특정부위를 포함하는 primer를 제작하고, 염기서열 분석을 통하여 유전자 구조를 해석한 결과 총 211bp 크기를 갖는 염기서열의 122번째에서 아데닌(A)${\leftrightarrow}$구아닌(G) 염기치환으로 발생한 단일염기다형(SNP) 부위를 발견하였다. 이들 단일염기다형 염기서열 부위를 PCR-SSCP(single-strand conformation polymorphism) 기법을 이용하여 분석한 결과 3종류의 SNP 유전자형(A/A, A/G 및 G/G)을 검출하였다. 이 가운데 A/G 유전자형이 한우의 근내지방도와 등지방두께와 고도의 유의적 연관성이 있다는 새로운 사실을 발견하였다. 따라서, 본 연구를 통해 개발된 한우 SCD 유전자의 특정한 단일 염기다형 표지인자는 한우의 연령 및 성별에 관계없이 육질이 우수한 고급육을 생산하는 우량 한우의 조기식별에 매우 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다.

• 대한화학회지
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• 제48권1호
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• pp.33-38
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• 2004

두경부 편평 세포암종(HNSCC: head and neck squamous cell carcinoma) 의 발생과 관련하여 p53 종양 억제 유전자 (tumor suppressor gene) 의 돌연변이는 높은 비율로 나타나는 것으로 보고 되고 있다. 단국대학교 병원에서 두경부 종양으로 진단 받고 수술 받은 환자의 조직 50개를 대상으로 p53 종양 억제 유전자의 exon 5-8 까지의 영역에서 DNA를 추출하여 PCR-SSCP(polymerase chain reaction single strand conformational polymorphism) 방법과 DHPLC(denaturing high performance liquid chromatography) 방법으로 p53체세포 돌연변이(somatic mutation)를 비교 분석하였다. 그 결과 SSCP 분석 방법은 16개(32%), DHPLC 분석 방법은 17개(34%) 를 검출하였고 그 중 SSCP와 DHPLC 분석 방법 모두 exon 8번에서 결실(deletion) 형태의 돌연변이를 확인하였으며 최종적으로 자동 염기 서열 분석기(automatic DNA sequencer) 를 통하여 모든 돌연변이를 확인하였다. DHPLC 분석방법이 SSCP 방법보다 분석 시간이나 노력이 덜 소모되며 보다 더 정확한 돌연변이 검출 방법임을 확인하였다.

• BMB Reports
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• 제36권4호
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• pp.399-402
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• 2003

Early detection of bladder cancer is particularly important since it dramatically affects the survival rates. However, neither urinary cytology nor tumor markers that are currently used are sensitive enough for the early detection of bladder cancer or recurrent disease. The ras genes are frequently mutated in cancer. In this study, we investigated the diagnostic potential of ras mutation analysis in urinary sediments of patients with bladder cancer using a single-strand conformation polymorphism analysis and polymerase chain reaction. Mutation in codon 12 of the H-ras gene was observed in 39% of the patients. Our results indicate that this approach may significantly improve diagnostic sensitivity in detecting bladder tumors.

• Journal of Microbiology
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• 제34권3호
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• pp.236-240
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• 1996

Rifampin is the most powerful drug for treating leprosy and tuberculosis today. It inhibits initiation and elongation of RNA transcription by binding to $\beta$-subunit of RNA polymerase, leading to kill mycobacteria. We isolated one variant strain of Mycobacterium leprae from 24 Korean leprosy patients who are less susceptible to rifampin or have suffered from relapse by polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) of the rpoB gene. Direct sequencing of the rpoB region of M. leprae variant revealed missense mutations which altered the amino acids sequenceof RpoB to Ser-464, Arg-465, Arg-467 and Ala-468. This is the first finding on rpoB gene mutation of M. leprae from Korean patients ; moreover the mutant type was found to be different from the previously reported cases in other countries.

• Asian-Australasian Journal of Animal Sciences
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• 제27권8호
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• pp.1075-1081
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• 2014

CRBP1 (cellular retinol binding protein 1) and CRBP3 (cellular retinol binding protein 3), are important components of the retinoid signaling pathway and take part in vitamin A absorption, transport and metabolism. Based on the role of vitamin A in chicken laying performance, we investigated the polymorphism of CRBP1 and CRBP3 genes in 349 chickens using single strand conformation polymorphism and DNA sequencing methods. Only one polymorphism was identified in the third intron of CRBP1, two polymorphisms were detected in CRBP3; they were located in the second intron and the third intron respectively. The association studies between these three SNPs and laying performance traits were performed in Erlang mountainous chicken. Notably, the SNP g.14604G>T of CRBP1 was shown to be significantly associated with body weight at first egg (BWFE), age at first egg (AFE), weight at first egg (WFE) and total number of eggs with 300 age (EN). The CRBP3 polymorphism g.934C>G was associated with AFE, and the g.1324A>G was associated with AFE and BWFE, but none of these polymorphisms were associated with egg quality traits. Haplotype combinations constructed on these two SNPs of CRBP3 gene were associated with BWFE and AFE. In particular, diplotype H2H2 had positive effect on AFE, BWFE, EN, and average egg-laying interval. We herein describe for the first time basic research on the polymorphism of chicken CRBP1 and CRBP3 genes that is predictive of genetic potential for laying performance in chicken.

• 생물정신의학
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• 제10권1호
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• pp.80-84
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• 2003

Objective : Bipolar disorder is known to have strong genetic background and cellular immune activation. Based on the hypothesis that abnormalities of normal inhibitory control of T cell immunity can contribute to the pathophysiology of bipolar disorder, we investigated the relationship between the first exon at position +49(A/G) polymorphism of cytotoxic T lymphocyte antigen 4(CTLA4) gene and bipolar disorder. Method : Among the Korean patients diagnosed as bipolar disorder according to DSM-IV, 90 patients without serious medical illness, neurologic illness, hormonal disorder, or concomitant mental illness were selected. The normal control group consisted of 149 age-and sex-matched subjects without current or past history of autoimmune diseases or mental disorder. DNA was extracted from whole blood and the exon 1 region of CTLA-4 gene was amplified by polymerase chain reaction. Gene typing was performed using single strand conformation polymorphism. Results : There were no significant differences in genotype frequencies of G/G, G/A, and A/A between the patients with bipolar disorder and the control group(48.9% vs 46.3%, 44.4% vs 39.6%, and 6.7% vs 14.1%, respectively). There were no significant differences in allelic frequencies of G and A between the patients with bipolar disorder and the control group(71.1% vs 66.1%, and 28.9% vs 33.9%, respectively). Conclusion : This study did not show the association of exon 1 polymorphism of CTLA-4 gene with bipolar disorder.

• Journal of Gastric Cancer
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• 제6권4호
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• pp.214-220
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• 2006

목적: 종양 억제, 세포 주기 조절 및 세포 고사의 기능과 연관 있는 유전자인 p53은 인간 종양에서 가장 흔히 발견되는 돌연변이 유전자로 알려져 있다. 위암에 있어서 p53의 돌연변이 정도와 생존율 등을 비교하여 각각의 상관관계와 예후 인자로써의 유용성에 대해 알아보고자 하였다. 방법: 1999년 3월부터 2001년 4월까지 경북대학교병원에서 위암으로 수술한 331명 환자의 조직을 이용하여, polymerase chain reaction single-strand conformation polymorphism 방법으로 p53 돌연변이를 확인하고, 환자의 임상 병리학적 인자와의 관계를 비교하였고, 환자의 생존율을 비교하였다. 결과: 전체 331명의 환자들의 조직 중 66예(19.9%)의 조직에서 p53 돌연변이가 관찰되었다. 이들 66예 중에서 exon 5에서 23예, exon 6에서 8예, exon 7에서 21예, exon 8에서 17예의 돌연변이를 보였는데, 이중 3예에서 2개의 exon에 돌연변이(exon 5와 exon 6, exon 6과 exon 7, exon 6과 exon 8)를 보였다 p53 돌연변이는 나이와 성별, 육안형, 병리학적 병기, 조직학적 분류, 종양의 위치에 따라서 차이는 없었으나, 장형 156예 중 36예(23.1%), 미만형 145예 중 19예(13.1%)로 유의한 차이가 있었고(P=0.007), p53 돌연변이에 따른 생존기간은 유의한 차이가 없었다(P=0.632). Exon 5는 장형(9.7%)에서 미만형(2.8%))보다 p53 돌연변이 빈도가 높았고(P=0.024), 림프절 전이가 있는 군에서 림프절 전이가 없는 군보다 p53 돌연변이의 빈도가 유의하게 높았다(25.0% vs 15.6%, P=0.034). 나머지 항목에서는 통계학적으로 유의한 차이가 없었다. 근치적 절제술을 시행한 예에서의 p53 돌연변이에 따른 생존율은 유의한 차이를 보이지 않았다(P=0.704). 결론: p53 돌연변이는 위암 환자의 예후인자로써 가치가 충분하지 않다.

• 한국산림과학회지
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• 제95권6호
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• pp.631-635
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• 2006

파이토플라스마 증폭 primer, R16F2n/R2를 이용하여 뽕나무 오갈병 파이토플라스마와 대추나무 빗자루병 파이토플라스마에 대하여 SSCP분석법을 이용하여 염기변이 분석을 하였다. 그 결과 뽕나무 및 대추나무 파이토플라스마는 약 1.2 kb PCR 산물을 이용하더라도 뚜렷한 밴드차이를 나타내었다. 유사한 SSCP밴드 패턴을 보이는 두 시료 간의 밴드형태를 뚜렷하게 구별하는 방법을 찾기 위하여 뽕나무 오갈병 파이토플라스마와 대추나무 빗자루병 파이토플라스마의 PCR산물을 혼합한 후 SSCP분석 결과, 전기영동상에서 대추나무 파이토플라스마와 뽕나무 파이토플라스마의 SSCP 밴드패턴 모두를 관찰할 수 있었다. 본 연구 결과, 기존에 약 600bp 크기로 한정된 것으로 알려진 SSCP 분석을 PCR 산물 1.2 kb을 이용하여 유사한 SSCP 밴드패턴을 보이는 두 시료간의 밴드형태를 두 시료의 PCR 산물을 혼합하여 SSCP분석함으로써 뚜렷하게 구별할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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• pp.1244-1249
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• 2014

Goose fatty liver is one of the most delicious and popular foods in the world, but there is no reliable genetic marker for the early selection and breeding of geese with good liver-producing potential. In our study, one hundred and twenty-four 78-day-old Landes geese bred in Shunda Landes goose breeding farm, Jiutai, Jilin, China were selected randomly. The fatty livers were sampled each week after overfeeding during a three week period. Polymerase chain reaction-single strand conformation polymorphism and DNA sequencing were used to identify single nucleotide polymorphisms (SNPs) of fatty acid synthase (FAS), which is an important enzyme involved in the synthesis of fat under both physiological and pathological conditions. Least-squares correlation was established between these SNPs and fatty liver weight, abdominal fat weight, and intestinal fat weight of the overfed Landes geese, respectively. The results showed that fatty liver weight of geese with EF and FF genotypes (amplified by primer P1) was significantly higher than that of the EE genotype (p<0.05), and liver weight of CD and DD genotypes (amplified by primer P2) was significantly higher than that of the CC genotype (p<0.05). Different genotype combinations showed different liver weights, and from highest to lowest were ABDD, DDEF, DDFF, DDEE, ABEF, ABFF, AADD, and CDEF. Further analysis of DNA sequencing showed that there were two SNPs within the 5' promoter region the FAS gene. The geese of EF and FF genotypes carried a change of T to C, and the geese of CD and DD genotypes carried a change of A to G. The changes of the bases could potentially influence the binding of some transcription factors to this region as to regulate FAS gene. To our knowledge, this is the first report of SNPs found within the 5' promoter region of the Landes goose FAS gene, and our data will provide an insight for early selection of geese for liver production.