소 모색관련 유전자 MC1R 의 RCR - RFLP Marker 를 이용한 한우육 판별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.4

The melanocortin 1 receptor(MClR) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat colour variations within several species including cattle. This study was performed to develop the identification technique of Hanwoo meat using MC1R gene associated with the coat colors of cattle. Alleles of the MC1R locus were detected by PCR-RFLP analysis and genotype frequency and DNA sequences of MC1R gene were compared among cattle breeds. Genomic DNA was extracted from meat or blood samples of five breeds including Hanwoo(n=200), Holstein(n=100), Angus(n=20), Hereford(n=20) and Charolais(n=20). The MC1R gene was used to amplify 739bp and 173bp of the bovine E-locus corresponding to positions 228-966 and 318-490, respectively, using the two specific primers. The amplified products were digested with Bse118 I or Msp I and Aci I enzymes, and DNA fragments were separated by gel electrophoresis for RFLP genotype analysis. Six genotypes, E^D/E^D E^D/E^+, E^D/e, E^+/E^+,E^+/e and e/e, controlled by three alleles E^D, E^+ and e were observed in MC1 locus. When the amplified DNA product(739bp) was digested with Bse118 I enzyme, Hanwoo meat showed a single band of 739bp, whereas two fragments of 531bp and 208bp were detected in Holstein meat and Angus breed, respectively. Also, in the RFLP patterns using Msp I enzyme, Hanwoo meat produced two fragments of 535bp and 174bp, while three fragments of 328bp, 207bp and 174bp were observed in Holstein meat and Angus breeds, respectively. Therefore, breed-specific RFLP markers showing distinct differences between these breeds were found by PCR-RFLP analysis. When the amplified DNA product(173bp) was digested with Aci I enzyme to classify subtype of E allele, the E^D allele produced three fragments of 97, 68 and 8bp, while the E^+ and d alleles produced two fragments of 173 and 8bp according to the Aci I recognition sequence. Among the six genotypes, two genotypes of E^+/e and e/e were observed in Hanwoo and their frequencies were 0.07 and 0.93, respectively. However, the E^D/ED and E^D/e genotypes were present in Holstein and E^D/E^D, E^D/E^+ and E^D/e genotypes in Angus breeds. Therefore, the E^+/e and e/e genotypes observed in Hanwoo and E^D/E^D, E^D/E^+ and E^D/e genotypes detected only in Holstein and Angus breeds may be useful as breed-specific DNA markers for distinguishing between Hanwoo meat and Holstein and Angus meats. When comparing MC1R sequences among Hanwoo, Holstein and Angus, a Gly → Val amino acid change due to a single base(G) deletion at colon 104 was found in Hanwoo. Consequently, breed specific RFLP genotypes of MC1R gene related to bovine coat colors could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

200 GeV/핵자 유황이온과 핵건판핵의 충돌에 의해 생성된 헬륨 파쇄핵의 극한파쇄 연구

김동철,송진섭,윤천실,정성헌,박인곤,김종오,김철수,김태연,이승희,조재희,천병구,김재률,김준원,김태익,박명렬,장한일,임인택 慶尙大學校 기초과학연구소 1992 基礎科學硏究所報 Vol.8 No.-

고에너지 중이온 원자핵과 핵건판의 충돌에서, 200GeV/핵자 유황이온에 의해 생성된 파쇄 헬륨핵(Z=2)의 실험실계의 방출각 분포는 표적핵에 무관한 회귀공식. dN=exp[a+k exp(η-y_b)]d[exp(η-y_b)]로 잘 표현된다. 여기에서 의사신속도 η=-ln[tan(θ/2)]이고, y_b는 실험실계의 입사입자(^32S)의 신속도이다. 이 공식에 의한 적합에서 k=-0.057±0.008로 얻어진다. 즉, 핵건판과 고에너지 중이온의 충돌에서 파쇄 헬륨핵의 exp(η-y_b)의 분포는 "극한파쇄" 현상을 잘 설명하고 있다. The angular distribution of emission angle θ of helium (Z=2) produced in the collisions of incident particles of 200 GeV/nucleon ^32S in nuclear emulsion is well expressed by dN=exp[a+k exp(η-y_b)]d[exp(η-y_b)] where the pseudorapidity is η=-ln[tan(θ/2)], the laboratory system primary rapidity is y_b, and k=-0.057+0.008. The shape of this frequency of occurrence distributions in terms of exp(η-y_b) attests to the validity of the concept of "limiting fragmentation" for helium projectile fragments produced in the projectile fragmentation regions of heavy ion collisions in nuclear emulsion.

PCR - SSCP 기법을 이용한 소 MC1R 유전자의 다형성 분석 및 한우육 감별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.1

The single-strand conformation polymorphism(SSCP) is a point mutation screening method based on changes in the secondary structure in a defined single-stranded DNA fragment caused by a change in sequence. This study was performed to develop the identification technique of Hanwoo meat using PCR-SSCP marker of MC1R gene associated with the coat colors of cattle. Hanwoo, Holstein and imported cattle breeds(Angus, Hereford and Charolais) were used for the detection of PCR-SSCP genotypes of MC1R gene. The specific primers were used to amplify a 138bp and a 390bp of the bovine E-locus(including point mutation) corresponding to positions 318-455 and 318-707, respectively, in the MC1R gene. For SSCP genotype analysis, the PCR products were denatured at 95℃ for 5min, loaded on a 12-15% nondenaturing polyacrylamide gel and detected by silver staining. When the PCR product of 138bp was separated by gel electrophoresis, two genotypes of E^+/e and e/e were detected only in Hanwoo meat, but E^D/E^D and E^D/e and E^D/E^D genotypes were observed in Holstein meat and Angus breed, respectively. Also, in the SSCP analysis using PCR product of 390bp, Hanwoo meat produced banding pattern of two fragments, while single fragment was observed in Holstein meat and Angus breed. Therefore, breed-specific SSCP markers showing distinct differences between Hanwoo and Holstein and Angus were found by PCR-SSCP analysis. This study shows that the PCR-SSCP technique is relatively simple, fast and a practical tool for determination of MC1R genotypes of cattle breeds when compared with PCR-RFLP method. Therefore, the PCR-SSCP markers of MC1R gene could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

TGF-β signaling plays an important role in resisting γ-irradiation

An, Y.S.,Kim, M.R.,Lee, S.S.,Lee, Y.S.,Chung, E.,Song, J.Y.,Lee, J.,Yi, J.Y. Academic Press 2013 Experimental cell research Vol.319 No.4

Transforming growth factor-β1 (TGF-β1) regulates various biological processes, including differentiation, bone remodeling and angiogenesis, and is particularly important as a regulator of homeostasis and cell growth in normal tissue. Interestingly, some studies have reported that TGF-β1 induces apoptosis through induction of specific genes, whereas others suggest that TGF-β1 inhibits apoptosis and facilitates cell survival. Resolving these discrepancies, which may reflect differences in cellular context, is an important research priority. Here, using the parental mink lung epithelial cell line, Mv1Lu, and its derivatives, R1B and DR26, lacking TGF-β receptors, we investigated the involvement of TGF-β signaling in the effects of γ-irradiation. We found that canonical TGF-β signaling played an important role in protecting cells from γ-irradiation. Introduction of functional TGF-β receptors or constitutively active Smads into R1B and DR26 cell lines reduced DNA fragmentation, Caspase-3 cleavage and γ-H2AX foci formation in γ-irradiated cells. Notably, we also found that de novo protein synthesis was required for the radio-resistant effects of TGF-β1. Our data thus indicate that TGF-β1 protected against γ-irradiation, decreasing DNA damage and reducing apoptosis, and thereby enhanced cell survival.

Comparison of helper component-protease RNA silencing suppression activity, subcellular localization, and aggregation of three Korean isolates of Turnip mosaic virus

Han, J. Y.,Chung, J.,Kim, J.,Seo, E. Y.,Kilcrease, J. P.,Bauchan, G. R.,Lim, S.,Hammond, J.,Lim, H. S. Springer Science + Business Media 2016 Virus genes Vol.52 No.4

<P>In 2014, we performed a nationwide survey in Korean radish fields to investigate the distribution and variability of Turnip mosaic virus (TuMV). Brassica rapa ssp. pekinensis sap-inoculated with three isolates of TuMV from infected radish tissue showed different symptom severities, whereas symptoms in Raphanus sativus were similar for each isolate. The helper component-protease (HC-Pro) genes of each isolate were sequenced, and phylogenetic analysis showed that the three Korean isolates were clustered into the basal-BR group. The HC-Pro proteins of these isolates were tested for their RNA silencing suppressor (VSR) activity and subcellular localization in Nicotiana benthamiana. A VSR assay by co-agroinfiltration of HC-Pro with soluble-modified GFP (smGFP) showed that HC-Pro of isolate R007 and R041 showed stronger VSR activity than R065. The HC-Pros showed 98.25 % amino acid identity, and weak VSR isolate (R065) has a single variant residue in the C-terminal domain associated with protease activity and self-interaction compared to isolates with strong VSR activity. Formation of large subcellular aggregates of GFP: HC-Pro fusion proteins in N. benthamiana was only observed for HC-Pro from isolates with strong VSR activity, suggesting that R065 'weak' HC-Pro may have diminished self-association; substitution of the variant C-terminal residue largely reversed the HC-Pro aggregation and silencing suppressor characteristics. The lack of correlation between VSR efficiency and induction of systemic necrosis (SN) suggests that differences in viral accumulation due to HC-Pro are not responsible for SN.</P>

표면탄성파 소자 응용을 위한 $LiTaO_3$ 단결정 성장

정대식,노용래,박병학,김유성,Chung, D.S.,Roh, Y.R.,Park, B.H.,Kim, Y.S. 한국결정성장학회 1994 한국결정성장학회지 Vol.4 No.2

표면탄성파 소자응요을 위하여 완전용융조성, 48.65 mole % $Li_2O에서 LiTaO_3(LT)$ 단결정(Y, X-axis)을 육성하였다. 육성된 $LiTaO_3(LT)$ 단결정의 SAW 소자성능을 확인하기 위하여 photolithography를 거쳐 기본적인 SAW filter를 제작하였다. Yamaju LT Y-cut 단결정 기판과 본 연구에서 제작된 RIST LT Y-cut 단결정 기판에서의 SAW특성결과를 비교하여 성능을 결정하였다. Yamaju LT 기판에서보다 본 연구에서 육성된 LT 단결정 기판에서 우수한 SAW 성능을 보였다. $LiTaO_3 (LT)$ single crystals (Y, X-axis) were grown from the congruent composition, 48.65 mole %, $Li_2O$ for SAW (Surface Acoustic Wave) applications. Basic SAW filters were fabricated on the RIST prepared LT wafers (Y-cut) using phtolithography. SAW filter performance was evaluated. The results were compared of the SAW characteristics between RIST prepared LT wafer (Y-cut) and commercial Yamaju wafer (Y-cut). The SAW filter prepared on the RIST grown LT wafer was shown better SAW performances than that of Yamaju wafer.

VNTR Marker 를 이용한 한우의 유전적 다양성 및 친자감별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.1

VNTR(Variable Number of tandem repeats) loci also known as hypervariable minisatellite loci are regions of DNA with highly polymorphisms in the number of tandemly repeated nucleotide sequences of around 25bp in length. This study was conducted to investigate allele frequencies and polymorphic information of BTGL1 VNTR locus and to check pedigree records by parentage testing and pedigree analysis of paternal half-sib families using VNTR markers in Korean cattle. The BTGL1 minisatellite locus was amplified by PCR and the DNA fragments were size-separated by non-denaturing polyacrylamide gels followed by silver staining. A total of 15 different alleles ranging in size from approximately 302 to 440bp were identified in the BTGL1 locus. The heterozygosity(H) and polymorphic information content(PIC) values were 0.877 and 0.828 in Korean cattle population, respectively. Mendelian segregation and codominant inheritance for the alleles of VNTR loci were confirmed in paternal half-sib families of Korean cattle. The allelic bands seen in the offsprings of each pedigree were always observed from either sire or dam. However, among the thirteen paternal half-sib families examined, the falsely recorded pedigrees were found in three families having non-paternal bands. Therefore, the relationships between parents and offspring in these three families can be excluded. The BTGL1 VNTR DNA polymorphisms could be used as genetic markers for routine parentage testing and pedigree analysis in Korean cattle.

임신기간중 혈장 Fibronectin 농도변화를 이용한 임신성 고혈압 발생의 예측 가능성에 관한 연구

정두용,최종현,최경남,김민철,승경록 건국대학교 의과학연구소 1995 건국의과학학술지 Vol.5 No.-

Preeclampsia is classically defined as the triad of pregnancy induced hypertension(PIH) with generalized edema and proteinuria, and constitutes a deadly triad of maternal death. Recent studies suggest that drugs affecting prostacyclin and thromboxane production may be able to affect the development of preeclampsia(PE). In order to use these agents effectively, a woman at high risk should be selected properly before the clinical illness manifests. As the pathophysiologic mechanism of PE., endothelial damage is thought to play a major role in these days. Many investigators have suggested that plasma fibronectin(FN), a good indicator of vascular endothelial damage, is markedly elevated at least 4 weeks before clinical manifestations of PE and can be used as a sensitive predictor of PIH. To assess the clinical utility of plasma FN in predicting PIH, plasma FN concentrations of pregnant women over 28 weeks of gestation who visited Kon-Kuk University Hospital from June 1, 1994 to February 28, 1995 were assayed. The results were as follows: among 108 pregnancy specimens, 70 were in the normal blood pressure group and 38 were PIH group: 20, transient hypertension subgroup(PH), 8, mild PE subgroup(PM), 10, severe PE subgroup(PS). There was statistical significance in the mean birthweight between normal blood pressure group(3,205±68g) and PS subgroup(2,455±315g) (p<0.05). The mean concentrations of plasma FN were highest in the PS subgroup(412±68.9ug/㎖) and lowest in the normal blood pressure group(244±8.7ug/㎖), that is stastically significant. Plasma FN concentration in the normal pregnancy period showed fluctuation: from 29 weeks of gestation to 36 weeks, elevation and after 36 weeks, decline. The changes in the mean concentrations of plasma FN after delivery were noted in both groups, but those in normal blood pressure group(254±13→227±10㎍/㎖) and PS(405±22→318±51㎍/㎖)were stastically significant(p<0.05). There was negative linear correlation between the platelet count and concentration of plasma FN in the PS group [platelets(Y)=-0.20192×FN(X)+262.23257 r=0.69] (p<0.05). There was no linear correlation between the birthweight and plasma FN concentration in normal blood pressure group and PIH group. The changes in the plasma FN concentration in the normal blood pressure group and PIH group before clinical manifestation were not statistically significant.

AFLP Marker를 이용한 한국 재래돼지의 유전적 다양성 및 품종식별

정의룡,김우태,김연수,이정구,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.6

본 연구는 한국재래돼지의 순수 혈통정립 및 유전자원 보존을 목적으로 AFLP 다형성을 분석하고 이를 유전적 표지인자로 이용하여 재래돼지의 유전적 특성을 분자 수준에서 규명하고 품종 집단내 유전적 변이성과 품종 특이적인 DNA marker를 탐색하며 동시에 타품종들과의 유전적 근연관계를 추정하기 위해 수행하였다. 13종류의 selective primer 조합형을 이용하여 분석한 결과 총 band의 수는 611개로 각 조합형 당 평균 47개의 band가 확인되었으며 이 가운데 다형적 band가 152개로 다형성 수준은 약24.5%였다. 한국 재래돼지의 다형율과 유전적 다양성 값은 각각 29.8%와 2.9로 개량종에 비해서 다소 높은 경향이었다. 한편, 13종류의 primer 조합형 가운데 E35/T38 및 E38/H13 primer 조합형에서 품종간의 차이를 나타내는 DNA marker가 확인되었다. 특히, 재래돼지에서 검출된 E35/T38 primer 조합형의 0.50kb, 0.25kb 및 0.38kb의 DNA band는 개량종과 명확히 구별되어 이들 품종 특이적 DNA band는 한국 재래돼지의 품종식별에 유용한 표지인자로 이용 가능할 것으로 기대된다. 품종간 유전적 근연관계에서 한국 재래돼지는 Hampshire종과 유전적 유사성이 가장 높은 것으로 추정되었다. 본 연구에서 검출한 한국 재래돼지의 AFLP 유전자 지문은 재래돼지 집단의 유전적 변이성 및 타 품종과의 근연관계 분석뿐만 아니라 경제형질과 연관된 marker 개발에 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다. For the purpose of genetic conversation and utilization of Korean native pig(KNP) as a valuable animal genetic resource, DNA polymorphisms of AFLP (amplified fragment length polymorphisms) as genetic markers were analyzed in KNP and foreign pig breeds (Landrace, Duroc, Hamphsire and Yorkshire). Using these AFLP markers, the genetic structure and characteristics of KNP population were analyzed at the DNA level and the genetic variability and diversity within and between breed population were evaluated. Breed-specific DNA marker for KNP was screened, and phylogenetic relationship within and among breeds were estimated. A total of 611 AFLP markers were amplified by 13 selective primer combinations, and the average number of bands per primer combination was 47.0. Among them 152 bands were polymorphic (24.5%). The rate of polymorphisms and genetic diversity values of KNP (29.8% and 2.90) was higher than these of other foregin breeds. E35/T38 and E38/H13 primer combinations produced AFLP banding patterns which clearly discriminated between KNP and other foreign pig breeds. Three bands(0.50kb, 0.25kb and 0.20kb) identified in E35/T38 and two bands(0.45kb and 0.38kb) identified in E38/H13 primer combinations were present in all of the KNP examined, but not present in the foreign pig breeds. Therefore, these two primer combinations could be used as breed-specific DNA markers for breed identification of KNP. In comparison of genetic distances among pig breeds, KNP was the most closely related to the Hampshire breed. AFLP fingerprints may be useful for genetic variability and relationships and development of breed-specific DNA markers in pig breeds.

한우 Mitochondrial DNA D-loop 영역의 염기서열 및 유전변이

정의룡,김우태,김연수,이정구,한상기 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.2

본 연구는 한우 mt DNA 염기서열 가운데 tRNA^Pro와 tRNA^Phe 유전자 사이에 위치하고 있는 D-loop 영역의 염기서열을 결정하고 한우의 염기변이 다형성을 분석하기 위하여 mt DNA 의 D-loop 영역내 404번부터 15061까지 1142bp 크기의 염기서열 부위의 특이적인 primer를 이용하여 PCR로 증폭하였다. 한우의 mt DNA내 D-loop 영역에서 단일염기의 치환 및 삽입에 의해 총 24개의 polymorphic site가 확인되었다. 한우 mt DNA D-loop 영역 가운데 16042∼16122번째에 위치하고 있는 영역의 염기치환율이 다른 염기서열 영역에 비하여 약 50%를 점유하고 있었으며 이들 polymorphic site에서 16055, 16230 및 16260 번의 염기치환은 한우에서만 검출된 새로운 염기변이로 확인되었다. 따라서, 한우 D-loop 영역내 특이적인 염기변이는 모계 및 세포질 DNA marker로서 한우품종을 특정하는데 활용할 수 있는 가능성을 시사하였다. Polymorphic site의 출현빈도는 169번째 염기가 81.3%로 가장 출현율이 높았고 다음으로 16302번과 16093번째는 56.3% 그리고 16042번와 16119번째 염기가 각각 50.0%와 43.8%의 치환율을 보였으며 나머지 14개 염기는 6.3%에서 25.0% 수준의 출현율을 나타냈다. 한우와 타 품종간의 유전적 근연관계를 추정한 결과 Bos taurus 계통의 유럽종과 유전적 유사도가 높은 것으로 추정되었고, Africa와 India의 Bos indicus 계통의 품종과는 상대적으로 근연관계가 먼 것으로 나타났다. 본 연구에서 검출한 mt DNA내 D-loop 영역의 염기서열 다형성은 한우집단의 유전적 변이성 추정과 모계유전 분석 및 나아가 경제적으로 중요한 형질들과의 연관성 분석에 유용하게 활용할 수 있을 것으로 기대된다. This study was performed to determine sequences of the mt DNA D-loop region, including tRNA^Pro and tRNA^Phe and to analysis sequence variation polymorphism in Korean cattle. The resulting sequencies were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify an 1142bp between nucleotides 15061 and 404 within the D-loop region of mt DNA using specific primers. Korean cattle showed 24 polymorphic sites by nucleotide substitutions and insertions of single base pairs. About 50% of polymorphic sites were found in positions 16042 to 16122 with the most variable region. Among these polymorphic sites, variations at 16055, 16230 and 16260 bp were detected as new sequence variants in Korean cattle. These specific polymorphic sites have not been reported in the Japanese black cattle and European cattle. Therefore, mt DNA variants in the D-loop region may be used as genetic markers for specifying Korean cattle. The frequencies of positions 169, 16302, 16093, 16042, 16119 with a high level of sequence polymorphism were 0.81, 0.56, 0.56, 0.50 and 0.43, respectively. In comparison of genetic distances. Korean cattle showed the more closely to European cattle as Bos taurus than Bos indicus such as African and India breeds. In conclusion, these mt DNA sequence polymorphisms in the D-loop region for Korean cattle may be useful for the analysis of cytoplasmic genetic variation and associations with economic important traits and genetic analysis of maternal lineage.