动物营养学报2019，Vol.31Issue(1)：266-273,8.DOI:10.3969/j.issn.1006-267x.2019.01.033
猪骨骼肌肌内脂滴分离和纯化方法的探究
Isolation and Purification of Intramuscular Lipid Droplets in Porcine Skeletal Muscle
摘要
Abstract
This experiment aimed to explore the establishment of an optimal method for the isolation and purification of intramuscular lipid droplets from porcine skeletal muscle, which would lay the foundation for further revealing the regulation mechanism of intramuscular fat formation.Longissimus dorsi muscle tissue from body weight of (120±2) kg"Duroc×Landrace ×Yorkshire"ternary hybrid finishing pigs was obtained after slaughter.The isolation of intramuscular lipid droplets of longissimus dorsi muscle was performed by ultracentrifugation.Explore the influence of different homogenization methods, temperature, buffer pH and phenylmethanesulfonyl fluoride (PMSF) on the isolation efficiency, and further determine the optimal centrifugal force and centrifugation time to purify lipid droplets.The results showed that the skeletal muscle was homogenized by a fully automatic tissue grinder, and the intramuscular lipid droplets were obtained by adding PMSF in 4 ℃, pH=7.4 buffer.With the purification conditions at 4 ℃, 230 000×g centrifugation with 30 min, the lipid droplets were the most abundant, the structure was complete, and the purity was the highest.In conclusion, the optimized isolation and purification method can obtain intramuscular lipid droplets of porcine skeletal muscles with complete morphology and high purity, which can open up new avenues to study intramuscular fat generation and the molecular regulation mechanism.关键词
猪/骨骼肌/肌内脂滴/超速离心/分离纯化Key words
porcine/skeletal muscle/intramuscular lipid droplets/ultracentrifugation/isolation and purification分类
农业科技引用本文复制引用
闫峻,马衍旋,穆淑琴,唐湘方,郑梓,李宁,刘陈龙,李千军,孙超..猪骨骼肌肌内脂滴分离和纯化方法的探究[J].动物营养学报,2019,31(1):266-273,8.基金项目
国家自然科学基金青年科学基金项目 (No.31402097) （No.31402097）
动物营养国家重点实验室开放基金 (7008) （7008）
天津市生猪产业体系创新团队资助 (IT-TPRS2017005) （IT-TPRS2017005）
中国博士后科学基金 (No.2014M561191) （No.2014M561191）
