摘要: |
600 g 冰冻羊垂体经过酸性溶液提取、硫酸铵的分级分离和 DEAE-纤维素柱层析得纯品(1),其得率为0.08g/kg,最后将部分纯品(1)过 Sephadex G-100分子筛,得到高纯的羊生长激素[纯品(2)]。提取的羊生长激素具有相当高的理化均一性,纯品(2):聚丙烯酰胺凝胶电泳呈现一条主带和二条酸性小带;聚丙烯酰胺等电聚焦电泳呈现二条带,其等电点分别为 pH7.35和 pH7.40,SDS—电泳呈现二条很难分开的带,其平均分子量为20 100道尔顿;用DNS-CI 法测得的 N-末端氨基酸是2个,丙氨酸和苯氨酸。纯品(1)理化均一性稍差:SDS—电泳较纯品(2)多出两条分子量低的带;等电聚焦电泳多出六条酸性小带。除此之外,其他理化性质同纯品(2)。用大白鼠胫骨实验测定生物活性,测得纯品(1)可使垂体大白鼠胫骨软骨盘的宽度相对增加79%。未对纯品(2)进行生物活性测定。用羊催乳素放射免疫药盒测定,纯品(1)和纯品(2)与羊催乳素均无交叉反应。 |
关键词: 羊垂体 生长激素 提取 鉴定 生物测定 |
DOI: |
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基金项目: |
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ISOLATION,CHARACTERIZATION OF BIOCHEMISTRY AND BIOASSAY IN OVINE GROWTH HORMONE |
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Abstract: |
Highly purified ovine growth hormone was obtained by extrac- tion of frozen pituitaries from acidic solution,by fractionation with methods of ammonium sulphate precipitation,chromatography on DEAE-cellulose (the average yield of about 0.08g/kg frozen pituitaries) and gel filtration through a column of Sephadex O-100.The protein obtained was homogeneous judged by the following items:polyacrylamide gel electrophoresis revealed two minor compo- nents accompanying a major anionic band,isoelectric focusing in po... |
Key words: ovine pituitary growth hormone isolation characterization bioassay |