보고서 정보
주관연구기관 |
국립축산과학원 National Institute of Animal Science |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2014-02 |
과제시작연도 |
2011 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201400011382 |
과제고유번호 |
1395021095 |
사업명 |
FTA대응 경쟁력향상기술개발 |
DB 구축일자 |
2014-06-28
|
DOI |
https://doi.org/10.23000/TRKO201400011382 |
초록
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Ⅳ. 연구개발결과
1. 비육기 SCD 및 FASN 유전자의 발현 양상 분석
○ 저 에너지 급여군에서 SCD유전자의 발현량이 고 에너지 급여 군에 비해 현저히 낮음을 확인
○ 향후 유전자의 발현에 따른 사료 성분 구성표 작성 등에 유의한 정보를 제공
○ SCD 유전자의 발현량과 지방산과의 상관관계분석
○ FASD6 대립유전자들이 Palmitoleic acid (C16:1n7), Vaccenic (18:ln7)및 Stearic acid (C18:0)의 분 산값들을 통계적인 유의수준에서 설명
○ SCD,
Ⅳ. 연구개발결과
1. 비육기 SCD 및 FASN 유전자의 발현 양상 분석
○ 저 에너지 급여군에서 SCD유전자의 발현량이 고 에너지 급여 군에 비해 현저히 낮음을 확인
○ 향후 유전자의 발현에 따른 사료 성분 구성표 작성 등에 유의한 정보를 제공
○ SCD 유전자의 발현량과 지방산과의 상관관계분석
○ FASD6 대립유전자들이 Palmitoleic acid (C16:1n7), Vaccenic (18:ln7)및 Stearic acid (C18:0)의 분 산값들을 통계적인 유의수준에서 설명
○ SCD, FASN, FADS6)에 대한 대립유전자 효과를 검증한 결과 상가적 대립유전자의 효과가 크게 작용함으로서 가축생산성 관련 유전자 마커로 활용
2. SCD와 FASN유전자형에 따른 한우 거세우의 에너지 급여 특성 구명
○ 한우시 보유축에 대한 불포화지방산 조성 관련 유전적 배경 구명 및 이를 이용한 마커 개발
○ 고품질 소비자 중심형 한우고기 생산체계 확립에 따르는 기초 자료 확보
3. 고 올레인산 한우고기 섭취 시 인체 내 건강기능성 구명
○ 고품질 소비자 중심형 한우고기 생산체계 확립에 따르는 기초 자료 확보
○ 건강기능성의 인체 시험 검증으로 우리나라 고유 축종인 한우고기 우수성 및 국제경쟁력 확보 가능한 과학적이고 실질적인 자료 제시
○ 지방산 조성에 따른 생체 생리학적 반응 및 기전 탐색으로 한우고기를 포함한 쇠고기의 건강유해성 논란 대응 자료
○ 한우고기의 맛을 결정하는 소비자 욕구와 시대적 요구변화에 능동적으로 대처하고, 향후 우리나라 한우농가의 쇠고기 생산 방향 제시
Abstract
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Characterization of differentially expressed SCD an FASN for the bovine carcasses according to the energy levels
It is an important fact that identifying genetic variations regarding quantitative traits (QTL) has been widely issued for domestic animal populations due to recent technologies of gen
Characterization of differentially expressed SCD an FASN for the bovine carcasses according to the energy levels
It is an important fact that identifying genetic variations regarding quantitative traits (QTL) has been widely issued for domestic animal populations due to recent technologies of genotyping and sequence analyses. According to the structural changes of genomic regions, genetic variants may explain variations of QTLs that were heavily subjected into the cattle breeding to maximize improvement of production rates. One of the fancy traits in animal breeding is to understand a mechanism of fat, which is a major issue in commercial meat industry regarding meat quality traits such as backfat thickness (BFT) and marbling scores (MAR) even though physiological features of fat related traits are still unclear. Furthermore, percentages of BFT and MAR influenced consumer preferences due to significant relationships between fat deposition and meat quality. Fatty acid composition also influenced a desirable flavor of meat products according to the percentage of monounsaturated fatty acids.
The purpose of this study was designed to discover genetic variants in the bovine fatty acid desaturase domain family member 6 (FADS6), fatty acid synthase (FASN), and Stearoyl-Co A desaturase (SCD) genes, to test associations with fatty acid composition as well as carcass traits such as backfat thickness (BFT) and marbling scores (MAR), and to identify expression patterns according to the administration of energy levels in diets. To amplify genomic and mRNA segments, a total of 12 primer pairs were designed from the genomic region based on nucleotide positions from 57,766,830 - 57,782,479 of the UCSC chromosome 19 (Baylor Btau_4.6.1/bosTau7). Sequence analyses detect 4 genetic variants in intron number 2 that is a huge non-coding region (approximately 10,890 bp). The FADS6 SNPs showed no significant departures from HWE except g.57772511C>T that did not have heterozygous genotypes. Genotypes of the g.57770744A>G and g.57772511C>T segments were significantly associated with fatty acid composition such as Vaccenic (18:ln7), Palmitoleic (16:ln7), and Stearic (18:0) acids. The analysis confirmed dominance and additive effects for the g.57770744A>G and g.57772511C>T segments, respectively. A positive (31.1%, P=0.003) and a negative (-36.5%, P<0.001) correlations between BFT and Linolenic acid (18:3n3) and between MAR and Eicosenoic acid (20:1n9) were observed, respectively. According to the feeding trails with different percentages of TDN levels, the present study confirmed significant changes of expression levels between lines, showing that the FADS6 gene was significantly expressed at least 2 folds with all FADS6 segments in the low line. Correlation analyses found that γ-Linoleic (18:3n6) showed a significant positive (R2=0.487, at the low line) and a negative (R2=0.5157, at the high line) correlations with expression levels of FADS6-M3.
The cluster of bovine fatty acid desaturase (FADS) gene consists of FADS1, FADS2, FADS3, and FADS6 that are key major enzymes in the fatty acid metabolism. To discover variants in FADS6, a total of 15 primer sets, which cover the whole FADS6 genomic regions, were designed. The analysis discovered 6 genetic variants that 3 SNPs in an intron 2 were genotyped with AS-PCR (Allele-Specific PCR) methods and 3 SNPs in an exon 6 were confirmed with RFLP (Restriction Fragment Length Polymorphism) procedures. Sequence alignments confirmed SNP positions located at 57770220, 57770489, 57771485, 57782205, 57782257, and 57782305, showing nucleotide substitutions as G/A, A/C, G/A, C/T, T/C, and C/T, respectively based on the reference sequences from UCSC (chromosome 19 : 57766830 - 57782480). The analysis revealed that palmitoleic acids (C16:1n7) was significantly associated with genotypes of g.57770220G>A, g.57770489A>C and g.57782305C>T while stearic acids (C18:0) showed significant associations with variants of g.57770489A>C segments.
Fatty acid composition (FAC) influences a desirable flavor of meat products according to the percentage of monounsaturated fatty acids (MUFA). The highest percentage of fatty acids in muscles has been known to oleic acid (18:1) that is believed to be a critical factor for a flavor of meat quality. In addition, saturated fatty acids (SFA) such as mirystic acid (14:0), palmitic acid (16:0), and stearic acid (18:0) influenced concentrations of lipoprotein cholesterol and fat hardness points. Considerable attention for FAC in meat products from cattle with high levels of SFA have been issued regarding nutritional supplements. Therefore, controlling concentrations of FAC in muscle tissues should be valuable according to the human health that need to optimize nutritional levels in beef diets. Dietary manipulations of FAC in order to decrease SFA levels and increase n-3 PUFA (poly unsaturated fatty acid) levels in ruminant meats have been widely used. A lipogenic enzyme SCD (stearoyl-CoA desaturase), which involves monounsaturated fatty acids (MUFA) biosynthesis, plays a critical role in muscle fat deposition. The SCD expression levels in response to different diet compositions should be responsible for influceing muscle FACs. The objective of this study was to identify differences of expression levels for SCD that was generated from different feeding trials with high and low energy levels. The FASN genotypes of the g.15532A>C locus were significantly associated with Linolenic (C18:3), and genotypes of the g.17924G>A locus were significantly associated with Palmitic (C16:0), Palmitoleic (C16:1), Oleic (C18:1), SFA, and UFA.
The experiment tended to show that long term administration of low and high energy diets may does not functionally either increase or decrease SFA and UFA. The interesting finding was that variances of concentrations for all FAC in the low energy diet group were higher than that of the high energy diet group except 18:2n6 and PUFA. However, statistical differences of variances for the average FACs between groups were not recognized. According to the variance analysis, the concentrations of FAC in the low group tended to have an approximate normal distribution while the high group showed a skewed patterns. Expression analysis of SCD revealed at least 2 fold higher expression levels in the high diet group, and especially, the SCD4 and SCD6 segments detected more than 10 folds higher expressions, while none of the SCD segments had high expression values in the low diet group. The results indicate that long term feeding with a higher proportion of nutritional levels lead to higher expression levels of the SCD gene.
In conclusion, excessive proportions of nutritional diets may have limitations for metabolic availabilities or functional restriction systems for the ruminal biohydrogenation between FAs when particular higher proportions of FAs are supplied. Therefore, the present results deliver a critical issue to the public that manipulations of FAC with higher nutritional levels may be oriented towards increases of expression levels of SCD, but not activities of other genes that functional elongate and desaturase FAs.
From the results, the analysis confirmed that the identified genetic variants would be useful molecular genetic markers to explain variations of fatty acid compositions as well as carcass traits for Hanwoo cattle. In addition, expression analysis presented a potential suggestion that genes are deferentially expressed according to the different administration of energy diets, but actual gene actions are not followed for physiological activities of genes. Therefore, it is required that animal breeders concern selecting sires lines for their population due to genetic effects that may not be involved in phenotypic values.
Genetic and gene expression analysis of SCD and FASN gene in Hanwoo carcass with different nutritional treatments
The aim of this experiment was to determine the effect of high energy diet on the level of SCD gene expressions and genotypes. High energy diet has been used for enhancing intramuscular adipose tissue in high quality beef cattle. High energy diet required not only an amount of concentrate on days of fattening periods but also induced cost for the management. We hypothesized that SCD expression is increased in beef cattle muscle tissue when fed a high energy diet and that this is related to SCD (CC, CT, TT) and FASN (GG, AG, AA) genotypes. A 2 x 3 factorial arrangement (High, Control, and CC, CT, TT) in a completely random design was used to feed 24 Hanwoo steers. Two steers were fed in same pen and 12 pens were used for treatment. Blood was drawn from each steers on every first week from 11 to 28 mo. Longissimus dorci (LD) muscle was collected within 10 min of harvest for analysis of mRNA SCD abundance. Overall ADG and feed efficiency were not different between high energy and control diet (P > 0.05). However, DMI were greater at control diet compare to high energy diet during final fattening periods (P < 0.05). Serum NEFA concentrations were greater in control diet than high energy diet group at final fattening periods (P < 0.05), but serum albumin, glucose, total protein, triglyceride, and phosphorus were greater in high energy group than those of control group (P < 0.05). Carcass traits or physco-chemical characteristics were not different between high and control fed Hanwoo steers. There was a negative correlation (R2=0.7868) between concentrations of stearic and palmitoleic acid. Palmitoleic acid was lowest in control-fed CT group and highest in control-fed TT group. Real-time quantitative PCR revealed that the mRNA content of SCD in muscle from high energy diet cattle increased (P < 0.05) as compared to the control group. The gene expression from muscle biopsy sample was indicated that high energy diet positively regulated mRNA level of PPARγ on day of final feeding period. These data indicated that high energy diet (+3% TDN) increased relative mRNA level of SCD, and these effects may be mediated monounsaturated fatty acid composition in Hanwoo LD muscle.
A study on investigation of healthy-function in man consuming oleic acid enriched Hanwoo beef
Ⅰ. 1st year (2011)
- Effect of ingestion of high and low MUFA beef on physiological metabolism of 20th to 30th men and women
Ingestion of hanwoo and imported beef did not significantly affect blood metabolites and lipid related index for 20th to 30th Korean men and women. This may be because the differences in oleic acid composition between Hanwoo and imported beef, in spite of higher oleic acid composition of Hanwoo beef compared to imported beef. Also, it may result from the insufficient refreshing period (3 wks) between two experimental periods for overcoming influence of the 1st period's ingestion in the 2nd period.
- Investigation of biometabolism of rat comsuming oil with different fatty acid composition
When rats comsumed oils with different fatty acid composition for 7 wks, daily body weight was significantly (p<0.05) affected to be 7.15g/d, 7.04, 6.74 and 6.62 for olive oil group, sunflower oil group, linseed oil group and control (no treated group), respectively. Albumin, glucose, total protein concentrations did not significantly (p>0.05) differ between the groups. HDL cholesterol and total cholesterol appeared to be highest for the sunflower oil group and lowest for control. GOT appeared to be highest for control (91.79 mg/dL) and lowest for the linseed oil group (55.06).
Ⅱ. 2nd year (2012)
- Effect of ingestion of high and low MUFA beef on physiological metabolism of 40th men and women
Ingestion of hanwoo and imported beef did not significantly affect blood metabolites and lipid related index for 40th Korean men and women. Hanwoo beef ingestion numerically increased HDL cholesterol for men and tended (p<0.10) to decrease HDL cholesterol for women, respectively. Also, numerical decrease in LDL cholesterol for men and women appeared when Hanwoo beef was consumed. Thus, present results indicates that the ingestion of Hanwoo beef, being high oleic acid composition, may be good or at least not negative for human health.
- Investigation of biometabolism of rat comsuming different levels of oleic acid intake
Concentrations of C18- fatty acids including oleic acid, C22:4 and DHA in plasma of rats consuming 4% or 8% oleic acid were dramatically higher than those of control in which rats consumed coconut oil. However, concentrations of HDL cholesterol, total cholesterol and trigylceride were not affected by the treatments.
Ⅲ. 3rd year (2013)
- Effect of ingestion of high and low MUFA beef on physiological metabolism of over 50th men and women
Ingestion of hanwoo and imported beef did not significantly affect blood metabolites and lipid related index for over 50th Korean men and women. But, LDL cholesterol, which has been known as 'bad cholesterol', was decreased by both beef ingestion.
Although HDL cholesterol concentration was not significantly (p>0.05) affected, ingestion of Hanwoo and imported beef numerically increased HDL cholesterol.
- Investigation of biometabolism of rat comsuming high and low MUFA beef
Compared with control including coconut oil, HDL cholesterol of rats consuming lipids of Hanwoo and imported beef was dramatically increased. Triglyceride of rats consuming Hanwoo beef lipid enriched oleic acids tended to be decreased. Also, ingestion of Hanwoo beef lipid was helpful for liver function related factors.
목차 Contents
- 표지 ... 1
- 제 출 문 ... 2
- 요 약 문 ... 3
- SUMMARY ... 6
- 목차 ... 11
- 제 1 장 서 론 ... 12
- 제 1 절 한우 SCD 및 FASN 유전자의 에너지 급여수준에 따른 유전자형 효과 및 도체내 발현양상 분석 ... 12
- 제 2 절 SCD와 FASN유전자형에 따른 한우 거세우의 에너지 급여 특성 구명 ... 12
- 제 3 절 고 올레인산 한우고기 섭취 시 인체 내 건강기능성 구명 ... 13
- 제 2 장 국내외 기술개발 현황 ... 14
- 제 1 절 한우 SCD 및 FASN 유전자의 에너지 급여수준에 따른 유전자형 효과 및 도체내 발현양상 분석 ... 14
- 제 2 절 SCD와 FASN유전자형에 따른 한우 거세우의 에너지 급여 특성 구명 ... 14
- 제 3 절 고 올레인산 한우고기 섭취 시 인체 내 건강기능성 구명 ... 15
- 제 3 장 연구개발 수행 내용 및 결과 ... 16
- 제 1 절 한우 SCD 및 FASN 유전자의 에너지 급여수준에 따른 유전자형 효과 및 도체내 발현양상 분석 ... 16
- 제 2 절 SCD와 FASN유전자형에 따른 한우 거세우의 에너지 급여 특성 구명 ... 81
- 제 3 절 고 올레인산 한우고기 섭취 시 인체 내 건강기능성 구명 ... 94
- 제 4 장 연구개발 목표 달성도 및 대외 기여도 ... 154
- 1절 : 목표대비 달성도 ... 154
- 2절 : 정량적 성과(논문게재, 특허출원, 기타) ... 154
- 제 5 장 연구개발결과의 활용계획 ... 158
- 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 158
- 제 7 장 기타 중요 변동사항 ... 159
- 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 159
- 제 9 장 참고문헌 ... 159
- 끝페이지 ... 169
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