초록 ▼

Ⅳ. 연구개발결과
1. 성장단계별 혈청 내 생리활성물질 지표 개발
○ 비육전기 혈중 생리활성물질은 ALB/T-PRO비가 높은 시험구에서 1+ 출현률 61%.
○ 비육후기 고급육 생산 거세한우의 혈중 포도당과 중성지방 농도가 높게 나타남
2. 한우 고급육 생산 조기 평가 방법 개발
○ 근육 위성세포 배양시스템을 구축
○ 한우 근육세포 특이적으로 발현되는 물질 확인 (TTR, FMOD)
○ 돼지 태반추출물을 이용한 지방축척 기작을 확립
3. 저지방 축적 부위의 근내 지방 축적율 향상 기술 개발

Ⅳ. 연구개발결과
1. 성장단계별 혈청 내 생리활성물질 지표 개발
○ 비육전기 혈중 생리활성물질은 ALB/T-PRO비가 높은 시험구에서 1+ 출현률 61%.
○ 비육후기 고급육 생산 거세한우의 혈중 포도당과 중성지방 농도가 높게 나타남
2. 한우 고급육 생산 조기 평가 방법 개발
○ 근육 위성세포 배양시스템을 구축
○ 한우 근육세포 특이적으로 발현되는 물질 확인 (TTR, FMOD)
○ 돼지 태반추출물을 이용한 지방축척 기작을 확립
3. 저지방 축적 부위의 근내 지방 축적율 향상 기술 개발
○ 지방세포로 이형분화유도 유전자 7종에 대한 발현 확인
○ 지방세포내의 특이발현 유전자인 PPARγ의 작용기전 확인
○ 지방분화 특이적으로 발생하는 microRNA에 대한 발현기작 확립

Abstract ▼

Development of an early prediction techniques for producing high-quality meat on diet and feedlot system
The aim of this project was to determine the effect of serum compositions predicted high quality meat. High energy diet has been used for enhancing marbling adipose tissue in high quality Hanw

Development of an early prediction techniques for producing high-quality meat on diet and feedlot system
The aim of this project was to determine the effect of serum compositions predicted high quality meat. High energy diet has been used for enhancing marbling adipose tissue in high quality Hanwoo beef. High energy diet required not only an amount of concentrate on days of fattening periods but also induced cost for the management. The selection technique for predicting high-quality beef cattle required a simple method and analysis. We hypothesized that a certain serum constitution or estimated breeding value (EBV) would be used to a prediction marker for high quality beef. A 2 x 2 factorial arrangement (High energy, control vs High EBV, Low EBV) in a completely random design was used to feed 26 Hanwoo steers.
Two steers were fed in same pen and 13 pens were used for treatment. Blood and body weight was collected from each steers on every months during early, middle, and final fattening periods. Longissimus dorci (LD) muscle and subcutaneous adipose tissue (SC) was collected within 10 min of harvest for analysis of adipogenic and myogenic mRNA levels. The LD samples were also collected after 24 hours chilled down for evaluating carcass grade or fatty acid composition. Over all ADG and feed efficiency were not different between high energy and control diet (P > 0.05). Dry matter intake was peaked by low EBV group from early and middle fattening periods(P < 0.05). Serum glucose and non-esterified fatty acid (NEFA) concentration were varied among treatments over all fattening periods (P < 0.05). There were no interaction between treatments at over all carcass traits (P > 0.05). Marbling score was numerically greater at high energy diet on high EBV groups than those of other groups. Total lipid percentage also tend to increase at high energy-high EBV groups. The percentage of palmitic acid was greater at high energy-high EBV groups than other groups from fatty acid compositions (P < 0.05).
The cystein and methionine percent of amino acid were greater at control diet-high EBV than other treatments in L. D. muscle compositions. Overall ADG and DMI were not different between high energy diet and control diet (P >0.05). Serum glucose and triglyceride concentrations were increased (P <0.05) by high EBV group from 22 to 28 month old. Serum NEFA concentrations were plateau at 24 months old high EBV group and steady reduced at 28 months old. (P <0.05). Marbling score and yield grade were not affected by high energy diet and EBV (P >0.05).
Real-time quantitative PCR revealed that the mRNA content of PPARγ and SCD of high EBV group increased (P < 0.05) as compared to low EBV group. The mRNA level of GLUT4 and MHC1 tend to increase when control diet fed at high EBV group. These data indicated that high energy diets increased serum glucose and triglyceride concentrations of high EBV-steers at final fattening period. Although there are no interaction between diet and EBV levels, adipogenic gene expressions in high EBV-steers were greater than those in low EBV-steers. The selection of EBV for meat quality affects serum level and adipogenic gene expression of feedlot Hanwoo steers.
Development of an early evaluation system by using serum to enhance meat quality.
The production costs and revenues in Hanwoo industry can be affected greatly by external environmental factors. Thus, according to the Korea-US FTA, the number of beef breeding farm income is expected to decrease, and the propagation in small farm are being rapidly dwindle by a combination of several factors such as burden and future operating costs. Therefore, development of early evaluation system is required to produce Hanwoo individuals and good quality meat. This work play an important role to increase competitiveness in Hanwoo industry.
The objective of this research works is the development of an early evaluation system to produce good quality meat through establishment of individual serum production, the development technique of serum character analysis, evaluation system to verify the ability of the muscle by serum, comparative analysis of experimental and feeding trial.
○ Serum securement and establishment of efficient production technology
○ Establishment of primary muscle satellite cell culture system
○ Observation of cell proliferation and differentiation using serum: Correlation of serum and meat grade was investigated by comparative analysis of cell culture with serum and feeding trial.
○ Gene expression analysis during muscle differentiation: TTR (transthyretin) and FMOD (fibromodulin) gene expression was analyzed during muscle differentiation with time and the relation of these two genes and calcium channels was observed by gene knock-down using shRNA.
○ Gene expression analysis during transdifferentiation: CD36 and FABP4 genes expression was observed in transdifferentiaton as adipocyte-like cell.
○ Lipid accumulation using a natural substance: lipid accumulation study was performed to identify accumulation causing factor with porcine placenta extract (PPSE) and muscle extract.
These results can generate physiological index and identify marker genes to increase meat quantity and quality. Non-preferred areas of meat can be converse as preferred region by improving lipid accumulation and this will be contribute to promote the consumption of non-preferred areas.
Development of an early prediction techniques for improvement of a fat deposition within low fat region of Hanwoo cattle In this study, we investigated the enhancement of intramuscular fat in nonpreferring meats of Hanwoo based on gene regulation mechanism. In 1st year, 7genes were selected to be involved in the transdifferentiation of myoblast C2C12 into adipocytes. Among them, PIK3R1 was selected and determined its transcriptional activation by PPARγ. Next, we investigated the transcriptional regulation of Mrap gene, which was isolated by ChIP on chip assay against PPARγ.
Suppression of Mrap expression induced delayed lipolysis in adipocytes. In 2nd year, we observed the direct interaction of PIK3R1 and PPARγ by ChIP assay. The upregulation of PPARγ-mediated PIK3R1 expression enhanced the insulin-stimulated AKT activation in 3T3-L1 cells. In addition, the ratio of lipid accumulation in 3T3-L1 cells was increased by gender specific serum. Serum derived from cows and steers induced more lipid accumulation than serum from bulls. In order to isolate gender specific microRNAs in bovine serum, microarray experiments were conducted. Selected miR-92a was highly distributed in steer's serum compared to bull's serum. The expression of miR-92a in adipocytes was regulated by PPARγ. In 3rd year, we investigated the transcriptional regulation of microRNAs by PPARγ.
Using bioinformatic analysis, promoter assay, and ChIP assay, adipogenesis-related microRNAs (miR-143, miR-107, and miR-17/92a) were transcriptionally activated by PPARγ. We had also isolated an additional microRNA by microarray assay. Isolat ed microRNA was miR-128, which was more distributed in steers than in bulls. The target gene of miR-128 was determined as Notch4 by reporter assay and ChIP assay. In addition, we presented miR-127 and miR-93 as candidate reference microRNAs in bovine serum for reliable expression data. It is necessary to choose suitable reference genes for qPCR analysis of serum. Our study will be contributed to investigate the molecular regulation of economic performance-related genes including the enhancement of intramuscular fat.

목차 Contents

• 표 지 ... 1
• 제출문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 5
• 목 차 ... 9
• 제 1 장 서 론 ... 10
• 제 1 절 영양 및 사양관리 시스템에 따른 혈청의 고급육 생산 능력 평가 방법 개발 ... 10
• 제 2 절 한우 혈청이 갖고 있는 고급육 생산 능력 조기 평가 방법 개발 ... 10
• 제 3 절 한우 저지방부위 근내 지방 축적률 향상 기술 개발 ... 12
• 제 2 장 국내외 기술개발 현황 ... 12
• 제 1 절 영양 및 사양관리 시스템에 따른 혈청의 고급육 생산 능력 평가 방법 개발 ... 12
• 제 2 절 한우 혈청이 갖고 있는 고급육 생산 능력 조기 평가 방법 개발 ... 13
• 제 3 절 한우 저지방부위 근내 지방 축적률 향상 기술 개발 ... 16
• 제 3 장 연구개발수행 내용 및 결과 ... 17
• 제 1 절 영양 및 사양관리 시스템에 따른 혈청의 고급육 생산 능력 평가 방법 개발 ... 17
• 제 2 절 한우 혈청이 갖고 있는 고급육 생산 능력 조기 평가 방법 개발 ... 42
• 제 3 절 한우 저지방부위 근내 지방 축적률 향상 기술 개발 ... 82
• 제 4 장 연구개발목표 달성도 및 대외기여도 ... 106
• 1절 : 목표대비 달성도 ... 106
• 2절 : 정량적 성과(논문게재, 특허출원, 기타) ... 107
• 제 5 장 연구개발결과의 활용계획 ... 109
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 109
• 제 7 장 기타 중요 변동사항 ... 109
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 109
• 제 9 장 참고문헌 ... 109
• 끝페이지 ... 114