• Journal of Animal Science and Technology
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• 제49권5호
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• pp.577-584
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• 2007

본 연구는 후대검정 한우 295두의 혈액으로부터 genomic DNA를 추출하여 PCR 방법에 의한 증폭과 염기서열 분석을 통하여 ADSF/resistin 유전자의 단일염기다형을 발굴하고 이들과 한우 육질관련형질과의 상관관계를 분석하기 위하여 실시하였다. 확보된 DNA로부터 염기서열을 결정한 결과 promoter와 4개의 exon영역에서는 SNP를 찾지 못하였으나 intron 영역에서 7개의 SNP를 발굴하였다. 발굴된 SNP의 출현 빈도는 0.027에서 0.16까지 그 차이가 많았다. 육질형질과의 상관분석에서 이들 SNP 중 intron 2에서 발굴된 764A ins 만이 근내지방도와 상관관계가 발견되었다(P<0.05). 근내지방도는 유전력이 매우 높기 때문에 이번에 발굴된 ADSF/resistin 유전자의 SNP 764A ins와 같이 유전표지인자를 이용하는 것이 근내 지방도의 개량을 위해 우수한 결과를 가지는 것으로 사료된다. 가축의 경제형질의 경우 다수의 유전자가 관여하기 때문에 한 개의 유전자를 이용한 가축의 선발 또는 개량에 이용한다는 것은 제한적일 수 있다. 따라서 다수의 관련 유전자를 이용한 다형현상과 경제형질과의 연관성 연구가 동반될 때 육질개선 및 가축개량에 실질적인 증대효과가 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.917-922
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• 2008

A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).

• Clinical and Experimental Pediatrics
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• 제51권7호
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• pp.771-774
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• 2008

가족성 저칼륨성 주기성 마비는 골격근에 존재하는 ion channel의 장애로 인해 저칼륨혈증과 연관되어 나타나는 주기성 이완성 마비를 보이는 드문 유전 질환이다. 이 질환의 발생에 관여하는 유전자로는 골격근육의 1q31-32 염색체에 위치하는 칼슘 채널과 나트륨 채널의 alpha subunit을 encoding하는 CACNA1S gene과 SCN4A gene이 밝혀져 있다. 국내에서도 소아의 가족성 저칼륨성 주기성 마비가 수 예 보고된 바 있지만, 유전자 분석을 통해 변이가 확인된 예는 드물다. 이에 우리는 CACNA1S의 Arg1239His 변이에 의한 저칼륨성 주기성 마비로 진단된 12세 환아의 증례를 보고하는 바이다. 이 변이는 현재까지 알려진 CACNA1S의 변이 중 비교적 흔한 것으로 알려져 있으나, 국내에서는 보고된 바가 없다. 저자들은 경구 acetazolamide와 칼륨 복용, 유발인자를 회피할 것을 교육함으로써 이 환아를 치료했으며, 현재 환아는 주기성 마비의 빈도와 중증도의 개선을 보이며 삶의 질 역시 향상되었다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7675-7679
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• 2013

Background: A missense mutation in exon 7 (R249S) of the p53 tumor suppressor gene is characteristic of aflatoxin B1 (AFB1) exposure. AFB1 is believed to have a synergistic effect on hepatitis virus B (HBV) carcinogenesis. However, results of studies comparing R249S prevalence among patients are conflicting. The aim of this study was to determine the prevalence of the R249S mutation in hepatocellular carcinoma (HCC) patients with or without positive HBsAg. Materials and Methods: Paraffin embedded liver tissues were obtained from 124 HCC patients who underwent liver resection and liver biopsy in King Chulalongkorn Memorial Hospital. Restriction fragment length polymorphism (RFLP) was utilized to detect the R249S mutation. Positive results were confirmed by direct sequencing. Results: Sixty four (52%) patients were positive for HBsAg and 18 (15%) were anti-HCV positive. 12 specimens tested positive by RFLP. Ten HCC patients (8.1%) were confirmed to be R249S positive by Sanger sequencing (AGG to AGT). Out of these 10, six were HBsAg positive, and out of the remaining 4, two were anti-HCV positive. The R249S prevalence among HCC patients with positive HBsAg was 9.4% compared to 6.7% for HBsAg negative samples. Patients with the R249S mutation were younger ($55{\pm}10$ vs $60{\pm}13$ year-old) and tended to have a more advanced Edmonson-Steiner grade of HCC, although differences did not reach statistical significance. Conclusions: Our study shows moderate prevalence of aflatoxin B1-related p53 mutation (R249S) in HCC with or without HBsAg. HBsAg positive status was not associated with R249S prevalence.

• Journal of Genetic Medicine
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• 제2권2호
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• pp.53-57
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• 1998

Spinal muscular atrophy (SMA) type I is a common severe autosomal recessive inherited neuromuscular disorder that has been mapped to chromosome 5q11.2-13.3. The survival motor neuron (SMN) gene, a candidate gene, is known to be deleted in 96% of patients with SMA type I. Presently, PCR and single strand conformation polymorphism (PCR-SSCP) analyses have been made possible for application to both archival slides and paraffin-embedded tissues. Archival materials represent valuable DNA resources for genetic diagnosis. We applied these methods for the identification of SMN gene of SMA type I in archival specimens for the prenatal diagnosis. In this study, we performed the prenatal diagnosis with chorionic villus sampling (CVS) cells on two women who had experienced neonatal death of SMA type I. DNA extraction was done from archival slide and tissue materials and PEP-PCR was performed using CVS cells. In order to identify common deletion region of SMN and neuronal apoptosis-inhibitory protein (NAIP) genes, cold PCR-SSCP and PCR-restriction site assay were carried out. Case 1 had deletions of the exons 7 and 8, and case 2 had exon 7 only on the telomeric SMN gene. Both cases were found to be normal on NAIP gene. These results were the same for both CVS and archival biopsied specimens. In both cases, the fetuses were, therefore, predicted to be at very high risk of being affected and the pregnancy were terminated. These data clearly demonstrate that archival slide and paraffin-embedded tissues can be a valuable source of DNA when the prenatal genetic diagnosis is needed in case any source for genetic analysis is not readily available due to previous death of the fetus or neonate.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7337-7341
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• 2014

Background: The physiological role of vitamin D extends beyond bone health and calcium-phosphate homeostasis to effects on cancer risk, mainly for colorectal cancer. Vitamin D may have an anticancer effect in colorectal cancer mediated by binding of the active form $1,25(OH)_2D$ to the vitamin D receptor (VDR). The Taq1 VDR gene polymorphism, a C-to-T base substitution (rs731236) in exon 9 may influence its expression and function. The aim of this study wass to determine the 25(OH)D vitamin D level and to investigate the association between circulating vitamin D level and Taq1VDR gene polymorphism among Jordanian colorectal cancer patients. Materials and Methods: This case control study enrolled ninety-three patients and one hundred and two healthy Jordanian volunteers from AL-Basheer Hospital/Amman (2012-2013). Ethical approval and signed consent forms were obtained from all participants before sample collection. 25(OH)D levels were determined by competitive immunoassay Elecsys (Roche Diagnostic, France). DNA was extracted (Promega, USA) and amplified by PCR followed by VDR Taq1 restriction enzyme digestion. The genotype distribution was evaluated by paired t-test and chi-square. Comparison between vitamin D levels among CRC and control were assessed by odds ratio with 95% confidence interval. Results: The vitamin D serum level was significantly lower among colorectal cancer patients (8.34 ng/ml) compared to the healthy control group (21.02ng/ml). Patients deficient in vitamin D (less than 10.0 ng/ml) had increased colorectal cancer risk 19.2 fold compared to control. Only 2.2% of CRC patients had optimal vitamin D compared to 23.5% among healthy control. TT, Tt and tt Taq1 genotype frequencies among CRC cases was 35.5%, 50.5% and 14% compared to 43.1%, 41.2% and 15.7% among healthy control; respectively. CRC patients had lower mean vitamin D level among TT ($8.91{\pm}4.31$) and Tt ($9.15{\pm}5.25$) genotypes compared to control ($21.3{\pm}8.31$) and ($19.3{\pm}7.68$); respectively. Conclusions: There is significant association between low 25(OH)D serum level and colorectal cancer risk. The VDRTaq1 polymorphism was associated with increased colorectal cancer risk among patient with VDRTaq1 TT and Tt genotypes. Understanding the functional mechanism of VDRTaq1 TT and Tt may provide a strategy for colorectal cancer prevention and treatment.

• 생명과학회지
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• 제12권2호
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• pp.173-181
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• 2002

북미산 S. commune UVMl-34의 A $\alpha$ 3 mating locus를 지니는 cosmid clone pSC13의 mating activity에 필요한 sequence 함유여부를 확인한 후 sequencing을 행하고 이에 대한 비교분석을 실시하였다. 그 결과 blast program을 사용한 전체 nucleotide 염기서열의 homology비교분석에서 남미산 1-71에 패하여 약 96%의 높은 homology를 나타내었으며, 1-71 A $\alpha$ 3 mating locus에서 모두 7개로 추정되는 exon의 염기서열의 비교실험에서도 거의 96% 이상에 이르는 homology를 나타내었다. 부분적으로는 AR (acidic rich region)에서 약 97%, HD (homeodomain)에서는 약 99%, BR (basic rich region)에서 약 97%, 그리고 Ser (serine rich regon)에서 약 95%의 높은 homology를 나타내어 지역간에 유전자상의 큰 변화는 나타나지 않는 것으로 확인되었다. 그리고 translated polypeptide sequence 를 이용하여 자생지가 다른 남미산 S. commune A $\alpha$ 3 mating locus 내의 Y-region은 비롯하여 다른 $A\alpha$ alleles내의 Y-region과 비교 분석한 결과 남미산에 대해 전체적으로 약 97%의 높은 homology를 나타내고 있지만 그 외 $A\alpha$ mating alllele gene의 Y-region과는 41~49%의 낮은 homology로 mating activity에 관여하고 있는 것으로 나타났다. 특히 mating에 있어 transcription regulator로 알려진 homeodomain에서는 약 98%의 homology가 나타남으로서 Z-region의 74%에 비하여 대륙간에 상당히 높은 유전자 보존상이 확인되었다. 또한 AR에서 97%, BR에서 100% 그리고 Ser에서도 98%의 상당히 높은 homology를 지니는 사실을 확인하였다. 이러한 결과로 보아 남미와 북미에 자생하는 같은 mating allele type간에는 상당히 높은 비율의 염기서열 보존이 이루어지고 있음을 알 수 있었으며, 비록 다른 $A\alpha$ alleles간의 비교 이지만 다른 mating alleles간의 약 50% homology와 비교할 때 보다 상당히 높은 결과로 보인다. 특히 homeodomain motif의 비교에서 Y1을 비롯한 다른 mating allele gene과도 85% 이상의 높은 homology를 나타내었으며 그 외 AR, BR, Ser 부위에서는 10~50%에 이르는 낮은 비율로 나타났다.

• 미생물학회지
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• 제54권4호
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• pp.309-319
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• 2018

이전의 연구에서 질소원이 존재하여도 페로몬을 유도하는 6개의 Schizosaccharomyces pombe 돌연변이체를 온도민감성 돌연변이체들의 저장고로부터 분리하였음이 보고된 바 있다. 본 연구에서는 이들 중 하나인 pws6 돌연변이체의 특성을 더 연구하였다. 이 돌연변이체는 영양물질에 특이적으로 페로몬 유도를 나타내었다. 즉 질소의 고갈은 없어도 M-factor 페로몬을 유도하였으나 탄소의 고갈이 없으면 유도되지 않았다. 이러한 결과는 pws6 돌연변이체가 질소 고갈을 전달하는 경로에 특이한 결함을 가지고 있음을 시사한다. 이 돌연변이체는 M-factor 페로몬뿐만 아니라 P-factor 페로몬도 온도에 민감한 양식으로 질소의 고갈 없이 유도함을 보여 주어 이 돌연변이체의 페로몬 유도는 세포 유형에 특이적이지 않음을 시사하였다. 이 돌연변이체의 온도 민감성 성장 결함의 상보적 보완에 의해 $pws6^+$ 유전자를 클로닝하여 8.1 kb, 3.3 kb, 그리고 4.8 kb 효모 DNA를 가진 3개의 플라스미드가 분리되었다. 이 플라스미드들은 pws6 돌연변이체의 성장 결함을 각각 100%, 70%, 그리고 10-20% 보완하였다. 또한 이 플라스미드들은 pws6 돌연변이체의 페로몬 유도 특성을 보완하는 능력을 가지고 있었으며 이는 돌연변이체의 성장 결함 보완 효율과 밀접한 연관성이 있음을 보여 주었다. 이들의 오픈 리딩 프레임을 성장 결함의 보완 효율과 비교하여 오픈 리딩 프레임 SPBC19C7.06이 pws6 돌연변이체의 온도 민감성 특성을 상보적으로 보완하는데 원인이 되는 리딩 프레임으로 결론 내렸다. 이 오픈 리딩 프레임은 prs1으로 명명되었으며 인트론이 없이 하나의 긴 엑손을 가지고 있는 추정된 prolyl tRNA synthetase를 암호화한다. 추정된 Prs1 단백질은 다른 종의 prolyl tRNA synthetase와 상당한 유사성을 보여 주었다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권12호
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• pp.1649-1659
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• 2012

Crosses between Korean and Landrace pigs have revealed a large quantitative trait loci (QTL) region for fat deposition in a region (89 cM) of porcine chromosome 4 (SSC4). To more finely map this QTL region and identify candidate genes for this trait, comparative mapping of pig and human chromosomes was performed in the present study. A region in the human genome that corresponds to the porcine QTL region was identified in HSA1q21. Furthermore, the LMNA gene, which is tightly associated with fat augmentation in humans, was localized to this region. Radiation hybrid (RH) mapping using a Sus scrofa RH panel localized LMNA to a region of 90.3 cM in the porcine genome, distinct from microsatellite marker S0214 (87.3 cM). Two-point analysis showed that LMNA was linked to S0214, SW1996, and S0073 on SSC4 with logarithm (base 10) of odds scores of 20.98, 17.78, and 16.73, respectively. To clone the porcine LMNA gene and to delineate the genomic structure and sequences, including the 3'untranslated region (UTR), rapid amplification of cDNA ends was performed. The coding sequence of porcine LMNA consisted of 1,719 bp, flanked by a 5'UTR and a 3'UTR. Two synonymous single nucleotide polymorphisms (SNPs) were identified in exons 3 and 7. Association tests showed that the SNP located in exon 3 (A193A) was significantly associated with weight at 30 wks (p<0.01) and crude fat content (p<0.05). This association suggests that SNPs located in LMNA could be used for marker-assisted selection in pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.627-632
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• 2007

FoxO1, FoxO3a and FoxO4 belong to the FoxO gene family, which play important roles in the PI3K/PKB pathway. In this study, we cloned the porcine FoxO1, FoxO3a and FoxO4 sequences and assigned them to SSC11p11-15, SSC1p13 and SSC xq13 using somatic cell hybrid panel (SCHP) and radiation hybrid panel (IMpRH). RT-PCR results showed that these three genes are expressed in multiple tissues. Sequencing of PCR products from different breeds identified a synonymous T/C polymorphism in exon 2 of FoxO3a. This FoxO3a single nucleotide polymorphism (SNP) can be detected by AvaII restriction enzyme. The allele frequencies of this SNP were investigated in Dahuabai, Meishan, Tongcheng, Yushan, Large White, and Duroc pigs. Association of the genotypes with growth and carcass traits showed that different genotypes of FoxO3a were associated with carcass length and backfat thickness between 6th and 7th ribs (BTR) and drip loss (p<0.05).