CAJ | 학술논문

　　选取宝石鲈肝胰脏作为样品，通过优化裂解配方及摸索裂解液与样品的最适比例，设计两个等电聚焦程序，建立和优化宝石鲈鱼肝胰脏蛋白质双向电泳实验方法，为宝石鲈肝胰脏内目标蛋白的研究提供参考。结果表明，采用裂解液配方：8 M Urea，2 M Thiourea，0.5%（W：V）CHAPS，40 mM Tris-Base，1%DTT，2%（W：V） Pharmalyte pH 3～10，1 mM PMSF；裂解液和样品液比为1∶3时裂解效果较好；等电聚焦程序I（S1：0～500 V，500 V·h；S2∶500 V，500 V·h；S3∶500～3500 V，10000 V·h；S4∶3500 V，50000 V·h，S5∶3500～5000 V，8000 V·h）具有最好的等电聚焦效果。
　　선취보석로간이장작위양품，통과우화렬해배방급모색렬해액여양품적최괄비례，설계량개등전취초정서，건립화우화보석로어간이장단백질쌍향전영실험방법，위보석로간이장내목표단백적연구제공삼고。결과표명，채용렬해액배방：8 M Urea，2 M Thiourea，0.5%（W：V）CHAPS，40 mM Tris-Base，1%DTT，2%（W：V） Pharmalyte pH 3～10，1 mM PMSF；렬해액화양품액비위1∶3시렬해효과교호；등전취초정서I（S1：0～500 V，500 V·h；S2∶500 V，500 V·h；S3∶500～3500 V，10000 V·h；S4∶3500 V，50000 V·h，S5∶3500～5000 V，8000 V·h）구유최호적등전취초효과。
Using the hepatopancreas of Jade perch as the experimental samples, the tissue lysate formula and ratio of lysis buffer volume to tissue weight, as well as the isoelectric focusing electrophoresis procedures were screened. The results were as follows:lysate formula I(8 M Urea, 2 M Thiourea, 0.5%(W∶V)of CHAPS, 40 mM Tris-Base, 1%DTT, 2%(W∶V)Pharmalyte pH 3-10, 1 mM PMSF)was the better. When the ratio Lysate volume to the sample weight was 1∶3, the lytic effect was the better. The electrophoretic program?(S1∶0-500 V, 500 V?h, S2∶500 V, 500 V?h, S3∶500-3 500 V, 10 000 V?h, S4∶3 500 V, 50 000 V?h. S5∶3 500-5 000 V, 8 000V?h)had the better isoelectric focused effect. Based on these results, a protocol of two-dimensional gel electrophoresis(2-DE)was set up. The optimized 2-DE techniques provided a basis for studying the proteome of Jade perch hepatopancreas.