摘要
胰岛素样生长因子Ⅰ（ＩＧＦ－Ⅰ）是一种多功能细胞增殖调控因子，它对许多疾病有较好的治疗作用．为了获得大量的ＩＧＦ－Ⅰ产品，在测定了ＩＧＦ－Ⅰ全长序列的基础上，构建了ＩＧＦ－Ⅰ表达载体ｐＢＶＩＧＦ，经热诱导表达后，ＳＤＳ－ＰＡＧＥ分析表明：含有重组表达质粒的菌株可表达出７．６ｋＤ的蛋白．研究了不同菌株对ＩＧＦ－Ⅰ表达的影响．ＩＧＦ－Ⅰ的表达水平可达１５ｍｇ／Ｌ．重组蛋白主要以包涵体形式存在，进行了初步纯化和复性研究，Ｗｅｓｔｅｒｎ印迹表明重组蛋白具有ＩＧＦ－Ⅰ的抗原性．并初步建立了ＩＧＦ－Ⅰ生物活性测定方法．
Insulin like growth factor Ⅰ(IGF Ⅰ)is a multifunctional cell regulating factor.It may be useful as a treatment of many diseases.After analysis of the complete sequence of IGF Ⅰ cDNA,IGF Ⅰ expression vector pBVIGF was constructed.SDS PAGE analysis suggested the bacteria containing the recombinant plasmid produce a novel protein of 7 6 kD as it was induced by raising the culture temperature.Different bacteria's influence on IGF Ⅰ expression was also taken on and the expression level of IGF Ⅰ reached to 15 mg/L.The recombinant protein was mainly recovered as inclusion body.IGF Ⅰ renaturation was on research after it was prelimitary purified.Western blotting indicated that recombinant protein could react with antibodies against anti IGF Ⅰ.IGF Ⅰ bioactivity assay was established.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1998年第1期47-51,共5页
Chinese Journal of Biochemistry and Molecular Biology
