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RISS 인기검색어
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RAPD - PCR 기법을 이용한 젖소의 DNA 다형분석과 유전적 특성에 관한 연구
정의룡(E . R . Chung),김우태(W . T . Kim),한상기(S . K . Han) 한국축산학회 1995 한국축산학회지 Vol.37 No.5
This study was carried out to detect DNA polymorphisms in domestic Holstein dairy cattle using random amplified polymotphic DNA(RAPD) analysis, based on the amplification of random DNA sequences with arbitrarily chosen primers by the PCR-technique, to analyze genetic variation and characteristics within the breed and to develop the breed-specific genetic markers by using these RAPD markers. Preliminary RAPD analysis was performed on genetic DNA samples from a total of 30 Holstein cows and 10 Korean native cattle as a reference breed using 40 different random primers. Of these, sixteen primers for further investigation were selected based on the number and frequency of the polymotphisms produced. Using 10 single-primers, a total of 78 RAPD markers were produced, seventy eight(`74.4%) bands of which were polymorphic, whereas twenty (25.6%) bands were monomorphic. The average bandsharing was 0.353 within the Holstein breed. Each of the primers generated RAPD banding patterns different in both size, number and intensity of amplified fragments. A specific primer (P-9) was found to be useful in the individual identification, resulting from the different DNA polymorphism among individuals. Two of the primers, P-5 and P-10, produced Holstein breedspecific RAPD markers when compared with those from the Korean native cattle. The RAPD profiles using polyacrylamide gel were different from those obtained in the agarose gel. The separation of the fragments with low molecular weight was more apparent in the polyacrylamide gel. The pairs of primers produced more or less DNA bands amplified than the single primers, depending on the composition of the two primer sequences combined.
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PCR - RFLP 기법을 이용한 젖소 유전적 개량을 위한 선발도구로서 β- lactoglobulin 좌위의 유전자형 분석
정의룡(E . R . Chung),김우태(W . T . Kim),한상기(S . K . Han) 한국축산학회 1994 한국축산학회지 Vol.36 No.6
Genotypes of the β-lactoglobulin(β-LG) locus as a generic marker linked to quantitative trait loci affecting traits of economic importance in dairy cattle were determined by the PCR-RFLP or AFLP method. Genomic DNA was prepared from blood of Holstein cows. The PCR was used to amplify a 262 by region between nucleotides 367 and 629 from exon IV to intron N of the bovine β-LG gene using forward primer(GTCCTTGTGCTGACACCGACTACA-3`) and reverse primer(CAGGACACCGGCTCCTGGTATATGA-3`). After amplification, PCR products were digested with Hae III restriction enzyme, and the fragments were separated by agarose gel electrophoresis for RFLP or AFLP analysis of β-LG locu s. RFLP specific for the β-LG A and B alleles were identified with the Hae III restriction enzyme. The β-LG AA genotypes produced two fragments of 109 and 153bp and the BB genotypes three fragments of 109, 79 and 74bp. The AB genotypes showed the intermediate pattern. Thus, PCR amplification and RFLP analysis was shown to be a rapid and sensitive method for the discrimination of β-LG genotypes directly at the DNA level in dairy cattle of any age or sex. Consequently, the PCR-RFLP or AFLP method presented in this study can be used as a valuable tool for early selection of A1 bulls and calves with the desirable β-LG gene or BB genotype affecting superior milk production traits for genetic improvement of dairy cattle.
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PCR - RFLP 기법을 이용해 젖소개량을 위한 유전적 표지로서 K- Casein 좌위의 유전자형 분석
정의룡(E . R . Chung),김우태(W . T . Kim),최석호(S . H . Choi),임태진(T . J . Rhim),한상기(S . K . Han) 한국축산학회 1995 한국축산학회지 Vol.37 No.1
Genotypes of K-casein(K-CN) locus as a genetic marker linked to quantitative trait loci affecting traits of economic importance in dairy cattle were determined by PCR-RFLP method. Genomic DNA was prepared from blood of Holstein cows. The PCR was used to amplify an 874 by region between nucleotides 10592 and 11466 from exon IV to intron IV of the bovine K-CN gene using sense primer(5`-GTGCTGAGTAGGTATCCTAG-3`) and antisense primer(5`GTAGAGTGCAACAACACTGG-3`). After amplification, PCR products were digested with four restriction enzymes, Hind III, Rsa I, Taq I, and Pst I, and the fragments were separated by agarose gel electrophoresis for RFLP analysis of K-CN locus. In addition to screening for the known Hind III and Rsa I restriction site polymorphisms of K-CN locus, we have found additional RFLPs specific for the K-CN A and B alleles in Taca I and Pst I enzymes. The amplified DNA product digested with each restriction enzyme generated specific RFLP pattern that allowed precise identification of K-CN AA, BB or AB genotypes. The K-CN genotypes determined for cows by the PCR-RFLP method agreed completely with the phenotypes obtained from milk samples of the same individuals. Thus, PCR amplification and RFLP analysis was shown to be a rapid and sensitive method for the discrimination of K-CN genotypes directly at the DNA level in dairy cattle of any age or sex. Consequently, the PCR-RFLP method presented in this study can be used as a valuable tool for early selection of AI bulls and calves with desirable K-CN B gene or K-CN BB genotype affecting superior milk production traits for genetic improvement of Holstein dairy cattle.
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재래한우의 보존을 위한 혈청 및 혈구단백질의 유전적 다형현상
한상기(S . K . Han),윤희섭(H . S . Yoon),정의룡(E . Y . Chung),신유철(Y . C . Shin),변희대(H . D . Byun) 한국축산학회 1995 한국축산학회지 Vol.37 No.1
Biochemical polymorphisms of five red cell and semen proteins, Hemoglobin(Hb), Transferrin(Tf), Post-transferrin 2(Ptf2), Post-albumin(Pa) and Albumin(Alb) as genetic markers in Korean cattle were analyzed by Starch and Polyacryamide gel electrophoresis and their phenotypes, genotypes and gene frequencies were estimated in order to analysis the genetic constitution of Korean native cattle population. In the Hemoglobin(Hb) locus four different phenotypes AA, AB, BB and CH were observed and assumed to be controlled by four different alleles designated Hb^A, Hb^B, Hb^C and Hb^H, and the Hb^H type was rare variant of Korean native cattle. The observed distribution of phenotypes were 73.37% for AA type, 23.37% for AB type. 2.72% for BB type and 0.54%r for CH type. Gene frequencies of Hb^A, Hb^B, Hb^C and Hb^H were 0.8505, 0.1440, 0.0027 and 0.0027. Semen Transfetrin(Tf) locus, 11 different phenotypes AA, AD₁, AD₂, AE, AH, D₁D₁, D₁D₂, D₁E, D₂H, D₂D₂, D₂E, EE and EH type were identified, which considered to be controlled by codominant alleles TF,^A Tf^D, Tf^D, Tf^E and Tf^H at a single locus. The frequencies of Tf genotypes AD₁, D₁E, D₁D₂, D₂E, AA, AE, D₁D₂, AD₂, D₁D₁, EE, AH, D₂H and EH were found to be 16.30, 13.33, 11.85, 10.37, 9.69, 8.15, 7.41, 9.63, 5.93, 4.44, 1.48, 0.74 and 0.01%, respectively. Gene frequencies of TF^A, Tf^(D1) Tf^(D2) and Tf^H were 0.2741, 0.2704, 0,2333, 0.2074 and 0.0148, respectively. And TfH gene were newly identified in Korean native cattle. Considering Post-transterrin 2 locus, three different phenotypess FF, FS and SS were identified, which considers to he controlled by two alleles Ptf^F and Ptf^S at a single autosomal locus. The frequencies of Rf genotypes FS, FF and SS were found to be 51.06. 36.88 and 12.06%n, respectively and gene frequencies of Ptf^F and Ptf^S were 0.6241 and 0.3759. In the Postalbumin(Pa) locus, three different phenotypes FF, FS and SS type were observed to be genetically controllled by Pa^F and Pa^S gene. And genotypes frequencies FS. FF amd SS type were 48.65, 36.(H and 1_5.32%, respectively. The gene frequencies of Pa^F and Pa^S were 0.6036 and 0.3964. The Albumin(Alb) locus were observed to lack any individual variation. Therefore, this locus were defined to be monomorphic. In comparison of genetic distance and dendogram calculated from the gene frequencies, close relationship was obtained between the Japanese cattle and the Korean cattle.
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제주재래마의 보존을 위한 생화학적 다형현상에 관한 연구 1 . 백혈구효소의 유전적 다형현상
한상기(S . K . Han),정의룡(E . R . Chung),신유철(Y . C . Shin) 한국축산학회 1993 한국축산학회지 Vol.35 No.5
Biochemical polymorphisms of five leucocytic enzymes, phosphoglucomutase(PGM), 6-phosphogluconate dehydrogenase (PGD), phosphohexose isomerase(PHI), mannose-6-phosphate isomerase(MPI) and glutamic oxaloacetate transaminase(GOT) were analyzed by starch gel electrophoresis and their phenotype, genotype and gene frequencies were estimated in order to analyze the genetic structure of Che Ju native horses. In the PGM locus, three different phenotypes FF, FS and SS were identified and assumed to be controlled by two autosomal alleles designated PGM^F and PGM^S. The genotype frequencies were 63.49% for SS, 25.40% for FS and 11.11% for FF. Gene frequencies of PGM^S and PGM^F were 0.7619 and 0.2381, respectively. In the PGD locus. three different phenotypes FF, FS and SS were recognized and assumed to be controlled by two autosomal codominant alleles designated PGD^F and PGD^S. The observed distributions of phenotype were 55.00% for SS, 37.00% for FS and 7.50% for FF. Gene frequencies of PGD^F and PGD^S were 0.2625 and 0.7375, respectively. In the PHI^F locus, two different phenotypes FI and II were observed and assumed to be controlled by two autosomal codominant alleles designated PHI^F and PHI^I, whereas the FF type was not recognized. The frequecies of PHI genotypes II and FI were found to be 76.62% and 23.38%, respectively and gene frequencies of PHI^F and PHI^I were 0.1169 and 0.8831, respectively. In the MPI locus, three different phenotypes AA, AB and BB were identified, which considered to be controlled by two codominant alleles MPI^A and MPI^B at a single autosomal locus. The distributions of phenotype BB, AB and AA were 67.92%, 20.75% and 11.32%, respectively and gene frequencies of MPI^B and MPI^A were 0.7830 and 0.2170, respectively. In the GOT locus, any individual variation was not found, therefore, this locus was defined to be monomorphic.
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제주재래마의 보존을 위한 혈청 단백질 및 효소의 유전적 다형현상
한상기(S . K . Han),정의룡(E . Y . Chung),신유철(Y . C . Shin),변희대(H . D . Byun) 한국축산학회 1995 한국축산학회지 Vol.37 No.1
Biochamical polymoiphisms of four serum proteins (Albumin(Alb), Vitamin D binding protein(Gc). Postalbumin(Pa) and Esterase(Es)) in Cheju native. horse were analyzed as genetic markers by electrophoresis and their phenotype, genotype and gene frequencies were estimated in order to determine the genetic structure of Cheju native horses. In the Albumin(Alb) locus, three different phenotypes FF, SS and FS were recognized and assumed to be controlled by two autosomal codominant alleles designated AIb^F and Alb^S. The observed distribution of phenotypes were 59.32% for FS type, 25.42% for SS type and 15.25% for FF type. Gene frequencies of Alb^F and Alb^S were 0.4491 and 0.5509, respectively. In the Vitamin D-binding protein(Gc) locus, two different phenotypes, FF and FS, were obsened and assumed to be controlled by two autosomal codominant alleles designated Gc^F and Gc^S whereas the SS type was not recognized. The frequencies of genotype FF, and SS, were found to be 92.19 and 7.81%, respectively and gene frequencies of Gc^F and Gc^S were 0.9609 and 0.0391. In the Post-albumin(Pa) locus, two different phenotypes, FF and FS, were identified, which were considered to be controlled by means of two codominant alleles and at a single autosomal locus, but the SS type was not recognized. The distribution of phenotypes were 98.44% for FF type, 1.04% for FS, and gene frequencies of Pa^F and Pa^S were 0.9948 and 0.00.52, respectivety. In the Esterase(Es) locus, four different phenotypes were identified and assumed to be controlled by alleles designated Es^F, Es^I, Es^S and Es^O. The observed distribution of phenotypes FI, II, IS, FS, FF, SS and 00 were 41.78, 30.18, 26.00, 18.00, 14.46, 5.60 and 1.43, respectively. Gene frequencies of Es^I, Es^F, Es^S and Es^Owre 0.4643, 0.3214, 0.2000 and 0.0143 respectively. In comparison of genetic distance and dendogram calculated from the gene frequencies, the most close relationship was obtained between the Cheju and Kiso horses and the lowest genetic similarity was obtained between the Cheju and Arabian horses.
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