배양 온도가 Porcine Liver Slice의 Protein Turnover에 미치는 영향 = Effect of the Cultiviting Temperature on the Protein Turnover in Porcine Liver Slice

다국어 초록 (Multilingual Abstract)

The study was investigated the effect of the cultivating temperature on the in vitro protein turnover in the swine liver slice. This study consists of two trials: protein degradation (first trial) and protein synthesis (second trial). In the first trial, swine liver slices incubated in Dulbecco's Modified Eagle's Medium(DMEM) at 32℃, 37℃, and 41℃ for 12 hours to testify the effects of the cultivating temperature on the in vitro protein degradation. The higher temperature went up, the more the degradation rates of protein in the swine liver slice increased. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), the protein concentrations in the ten bands (MW: 116.5kDa, 51.1kDa, 86.0kDa, 78.5 kDa, 34.9kDa, 89.9kDa, 60.1kDa, 40.8kDa, 28.9kDa, and 19.6kDa) were analyzed by the densitometer. The protein concentrations of the 116.5 kd and 51.1 kd bands decreased as the cultivating temperature went up. The protein concentrations of 86.0kDa, 78.5kDa, and 34.9kDa bands were not affected by the cultivating temperature. The protein concentrations of the 89.9kDa and 60.1kDa bands were not different between at 32℃ and 37℃, but was remarkably lower at 41℃ than at 32℃ and 37℃. The protein concentrations of the 40.8kDa, 28.9kDa, and 19.6kd bands were similar at 37℃ and 41℃, but reduced at 37℃ and 41℃ compared to 32℃. In the second trial, swine liver slices incubated in DMEM containing fetal bovine serum (FBS) and 35S-methionine at 32℃, 37℃, and 41℃ for 12 hours to prove the effects of the cultivating temperature on the in vitro protein synthesis. The rates of protein synthesis in swine liver slice were the highest at 37℃ and the lowest at 32℃. After SDS-PAGE, the amounts of newly synthesized proteins in 109kDa, 100kDa, 91kDa, 75kDa, 63kDa, 52kDa, 45kDa, 37kDa, 30kDa and 21kDa bands were analyzed by the autoradiograph and the densitometer. The amounts of newly synthesized proteins in all bands were also the highest at 37℃ and the lowest at 32℃. In conclusion, the results of these studies suggested that in swine liver slice system, the rates of protein degradation increase as temperature go up and the rates of protein synthesis are the highest at normal body temperatures.

목차 (Table of Contents)

• Ⅰ. ABSTRACT 1
• Ⅱ. 서론 4
• Ⅲ. 연구사 6
• 1. 고온 스트레스가 가축의 생산성에 미치는 영향 6
• 1.1) 반추동물 6
• 1.2) 가금류 9
• 1.3) 돼지 12
• 2. 고온 스트레스 시 돼지의 생리적 변화 14
• 3. 고온 스트레스 시 protein turnover에 대한 연구 16
• Ⅳ. 재료 및 방법 18
• 1. 공시동물 18
• 2. 돼지 간의 slice에서 단백질 양과 단백질 합성측정 18
• 2.1) 돼지 간의 slice 준비 18
• 2.2) 돼지 간의 slice의 배양 및 세포질 단백질의 분석 19
• 2.3) 돼지 간의 조직으로부터 단백질 합성 24
• 3. 전기영동 및 Radiography 25
• Ⅴ. 결과 및 고찰 26
• 1. 배양 온도가 돼지 liver slice의 단백질 농도에 미치는 영향 26
• 2. 배양 온도가 돼지 liver slice의 단백질 합성에 미치는 영향 34
• Ⅵ. 적요 40
• Ⅶ. 참고 문헌 42