선택한 단어 수는 입니다.
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선택한 단어 수는 30입니다.
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가설 설정
- The mtDNA sequence reported in GenBank did not have Msp I restriction sites on the region of cytochrome oxidase complex I, II and III. It may be explained by hypotheses that either Hanwoo has different genetic composition comparing with Bos Taurus cattle or the Msp I restriction site is an SNP (single nucleotide polymorphism). Even though Hanwoo is in a phylogenetic line of Bos Taurus for the evolutional relationship, some part of genetic constitution may differ.
- It may be explained by hypotheses that either Hanwoo has different genetic composition comparing with Bos Taurus cattle or the Msp Ⅰ restriction site is an SNP (single nucleotide polymorphism).
제안 방법
- Least squares means and standard errors were determined for all measurements with a model including fixed effects of the cytochrome oxidase subunit genotypes, parity, and castration, and a covariate for age of animal. Analysis of variance was conducted using Statistical Analysis System (SAS) general linear models (GLM) procedures, and least squares means were compared using Fisher's least significant difference test (SAS, 1985) with a comparison error rate of 0.
- Polymerase chain reaction (PCR) was conducted with a final volume of 20 μl, including 2 μl of 10 x reaction buffer (10 mM Tris, pH 8.3, 50 mM KCl, 0.1% Triton X-100, 1.5 mM MgCl2), 10 μM dNTP, 10 pM of each primer, 50 ng of genomic DNA, and two units of Taq DNA polymerase.
- Restriction fragment length polymorphism patterns for Hanwoo mtDNA using 8 restriction enzymes (M: size marker, lane 1 and 2: mt5 digested by Hinf I, lane 3 and 4: mt6 digested by Rsa I, lane 5 and 6: mt6 digested by Msp I, lane 7 and 8: mt6 digested by Bgl Ⅱ, lane 9 and 10: mt7 digested by EcoR I, lane 11 and 12: mt8 digested by Hinf I, lane 13 and 14: mt8 digested by Rsa I, lane 15 and 16: mt9 digested by Pvu Ⅱ).
- Eight out of 20 restriction enzymes (Bgl I, Bam HI, Hind III, Hae II, Xho I, Kpn I, Apa I and Sma I) were found no restriction cleavage sites in the region of cytochrome oxidase subunit of Hanwoo mtDNA. Restriction sites were simulated using mtDNA sequences from GenBank to compare the differences of sequences and restriction sites for our findings. The results were corresponding to the results from simulated restriction sites using the sequences from GenBank.
대상 데이터
- Genetic variants of mtDNA were revealed by restriction endonuclease analysis. A total of 20 restriction enzymes (Alu I, Hae III, Rsa I, Sca I, Msp I, Bgl I, Bgl II, Hinf I, Bam HI, Hind III, Pst I, Pvu II, Hae II, Xho I, Hph I, Kpn I, Apa I, EcoR I, Taq I and Sma I) were used. All the restriction enzymes had more than 2 restriction cleavage sites for the region of cytochrome oxidase subunit in Hanwoo mtDNA (Figure 1).
- A total of 20 restriction enzymes (Alu I, Hae Ⅲ, Rsa I, Sca I, Msp I, Bgl I, Bgl Ⅱ, Hinf I, Bam HI, Hind Ⅲ, Pst I, Pvu Ⅱ, Hae Ⅱ, Xho I, Hph I, Kpn I, Apa I, EcoR I, Taq Ⅰ and Sma I) were used.
- Forty-one different types of restriction enzymes having 214 cleavage sites in the region of cytochrome oxidase were simulated. All the restriction cleavage sites detected in this experiment were belonged to the 214 of simulated cleavage sites, which were generated by computer programs except for Msp I restriction site. The mtDNA sequence reported in GenBank did not have Msp I restriction sites on the region of cytochrome oxidase complex I, II and III.
- All the restriction cleavage sites detected in this experiment were belonged to the 214 of simulated cleavage sites, which were generated by computer programs except for Msp Ⅰ restriction site.
- Six microliter aliquots from each PCR reaction mixture were used for restriction endonuclease digestion using 2 units of enzymes under the conditions suggested by the manufacturers. The enzyme used in this experiment were: Alu I, Hae III, Rsa I, Sca I, Msp I, Bgl I, Bgl II, HinfI, Bam HI, Hind III, Pst I, Pvu II, Hae II, Xho I, Hph I, Kpn I, Apa I, Eco RI, Sma I and Taq I (Promega). These restriction enzymes showed sufficient activity to be used directly in the PCR mixture.
- The enzyme used in this experiment were: Alu I, Hae Ⅲ, Rsa I, Sca I, Msp I, Bgl I, Bgl Ⅱ, Hinf I, Bam HI, Hind Ⅲ, Pst I, Pvu Ⅱ, Hae Ⅱ, Xho I, Hph I, Kpn I, Apa I, Eco RI, Sma Ⅰ and Taq Ⅰ (Promega).
- Two hundred thirty one Korean native steers, which were part of the 33rd progeny test in 2002 and 143 bulls, were used from Hanwoo Experiment Station ofthe National Livestock Research Institute (NLRI). The cattle were fed a postweaning corn and soybean meal diet, which was formulated to meet NRC (1984) requirements for growing beef cattle.
- Two hundred thirty one Korean native steers, which were part of the 33rd progeny test in 2002 and 143 bulls, were used from Hanwoo Experiment Station ofthe National Livestock Research Institute (NLRI). The cattle were fed a postweaning corn and soybean meal diet, which was formulated to meet NRC (1984) requirements for growing beef cattle.
데이터처리
- Analysis of variance was conducted using Statistical Analysis System (SAS) general linear models (GLM) procedures, and least squares means were compared using Fisher's least significant difference test (SAS, 1985) with a comparison error rate of 0.05.
- Least squares means and standard errors were determined for all measurements with a model including fixed effects of the cytochrome oxidase subunit genotypes, parity, and castration, and a covariate for age of animal. Analysis of variance was conducted using Statistical Analysis System (SAS) general linear models (GLM) procedures, and least squares means were compared using Fisher's least significant difference test (SAS, 1985) with a comparison error rate of 0.05. A total of eight mtDNA segments were genotyped, and all allele effect for the segments was analyzed separately.
성능/효과
- All the restriction enzymes had more than 2 restriction cleavage sites for the region of cytochrome oxidase subunit in Hanwoo mtDNA (Figure 1). Eight out of 20 restriction enzymes (Bgl I, Bam HI, Hind III, Hae II, Xho I, Kpn I, Apa I and Sma I) were found no restriction cleavage sites in the region of cytochrome oxidase subunit of Hanwoo mtDNA. Restriction sites were simulated using mtDNA sequences from GenBank to compare the differences of sequences and restriction sites for our findings.
- Eight out of 20 restriction enzymes (Bgl I, Bam HI, Hind Ⅲ, Hae Ⅱ, Xho I, Kpn I, Apa Ⅰ and Sma I) were found no restriction cleavage sites in the region of cytochrome oxidase subunit of Hanwoo mtDNA.
- From our results, mtDNA polymorphisms were detected by restriction enzymes (Pst I, Pvu Ⅱ, Rsa I, Eco RI, Bgl Ⅱ and Msp I), showing genetic differences in the region of cytochrome oxidase subunit I, Ⅱ and Ⅲ in Hanwoo.
- Therefore, we may expect that significant genetic effects on growth traits may be observed in the late age stages. In this study, genotypic effects on weight were found in the late age stages from WT15 and WT18. However, it is still unclear that these results are from either hypothesis of mtDNA mutation correlated to animal aging and development process or statistical differences of different rate of gene expression for individuals in the late growing stages.
후속연구
- Even though some preliminary results for mtDNA effects on important traits, extensive researches were not taken because of shortage for the structured data set, which is materially related Hanwoo population in several generations. Therefore, to better understand the effects of mtDNA genotype on growing stages, it may be necessary further investigation with well unequivocally assigned Hanwoo population.
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