The he was strongly inhibited by sbti and apmsf , and very sensitive to pmsf , lbti , and tlck , while not sensitive to chymostatin , bestatin , leupeptin , tpck , pepstatin , nem and iam . all these results imply that brine shrimp he was most probably a trypsin - type serine protease . the he could be strongly inhibited by edta in a dose - dependent manner , and 50 mmol / l edta exhibited more than 56 . 5 % inhibition 对孵化酶纯化样品进行生化性质和酶性质分析发现，卤虫孵化酶的最适反应温度约为40 ，最适ph为8 . 5左右；该孵化酶对p - apmsf 、 sbti极为敏感，对pmsf 、 lbti和tlck也非常敏感，但对chymostatin 、 leupeptin 、 pepstatin 、 bestatin 、 tpck 、 nem和iam不敏感，表明该酶极可能是一种属于胰蛋白酶类型的丝氨酸蛋白酶。
2.
The enzyme activity could be completely inhibited by pmsf and dfp , and partially inhibited by pepstatin a , edta and egta . leupeptin , benzamidine and hydrochloride aprotin had no influence on the dhap activity . ca2 * , mg2 + and na * could slightly increase , but cu2 + and zn2 + slightly inhibited protease activity 此结果表明， dhap为一种丝氨酸蛋白酶，其酶活性的发挥需要某些金属离子的参与，如na + ， c扩+ ， m扩+等金属离子能提高dhap的活性，但c了+与z矛十能抑制dhap的部分活性。
3.
Camp , pka inhibitor peptide ( pki ) , kemptide , histone h1 ( type iii - s ) , polyvinyl alcohol , na3vo4 , naf , b - glycerophosphate , p - nitrophenylphosphate , 3 - [ n - morpholino ] - propanesulfonic acid ( mops ) , genistein , ml - 9 , leupeptin , aprotonin , pepstatin , chymostatin , trypsin - chymotrypsin inhibitor were purchased from sigma chemical co . [ r - 32 p ] atp was obtained from yahui biomedical enginnering co 实验材料昆明系小白鼠，中国医科大学实验动物部提供。测活用试剂除nacl山inni为国产分析纯对rr为amesa产品， y乍jatp为北京亚辉生物医学工程公司产品外，余者均为sipoa产品。
4.
Western blot analysis we treated 200 fertilized eggs in a series of steps including lysing them in l0ul of lysis buffer ( 50mm tris - hcl ph 7 . 5 , 250mm nacl , 5mm edta , imm dtt , 0 . 1 % triton , 50mm sodium orthovanadate , looug / mg pmsf and tpck , 50ug / ml tlck , 1 ug / ml leupeptin pepstatin and aprotinin ) , frozing them below - 70 , and then thawing them at room temperature for 10 min 2 . westem印迹将处理组及对照组小鼠一细胞g2期受精卵各200个取出，转移到ep - pendoff管中， 5000甲m离心4分钟，弃去上清，加入ro川粉碎缓冲液，充分振荡混匀，在液氮中经过2一3次的冻融循环，迫使卵细胞裂解，加人等量的样品缓冲液沸水中煮5分钟，用于westem印迹分析。
