You Position: Home > Paper
Soluble expression of human carboxypeptidase A1 active center in pichia yeast
( views:228, downloads:4 )
- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 5
- DOI:
- 10.3321/j.issn:1000-2790.2006.05.002
- Key Word:
- 羧肽酶类A;活性中心;毕赤酵母;基因表达
Abstract： 目的:应用毕赤酵母表达系统表达人羧肽酶A1(carboxypeptidase A1,CPA1)活性中心蛋白.方法:从pGEX-CPA1重组质粒中扩增出CPA1活性中心基因,亚克隆入酵母表达载体pPIC9K,酵母重组表达载体pPIC9K-CPA1active center电转化入毕赤酵母SMD1168,并进行营养缺陷筛选和G418抗性筛选,对高拷贝整合的重组酵母表达菌株诱导表达.结果:构建得到酵母融合重组表达载体pPIC9K-CPA1active center,经G418浓度梯度筛选得到串联整合16个重组表达载体的重组酵母表达菌株,诱导表达后得到CPA1活性中心蛋白.结论:在毕赤酵母中成功表达了CPA1活性中心,为进一步研究CPA1活性中心在抗体导向酶-前体药物疗法中的应用打下了基础.
- [1]岳乔红,于文彬,郝晓柯,等.人羧肽酶A1活性中心基因的克隆及原核表达[J].2003,(23).
- [2]Joshi L,St Leger-RJ.Cloning, expression, and substrate specificity of MeCPA, a zinc carboxypeptidase that is secreted into infected tissues by the fungal entomopathogen Metarhizium anisopliae.[J].1999,274(14).
- [3]Segura S,Feng Y,Vendrell J,etc.Identification and characterization of three members of the human metallocarboxypeptidase gene family.[J].2002,277(17).
- [4]J L, Cereghino,J M, Cregg.Heterologous protein expression in the methylotrophic yeast Pichia pastoris.[J].2000,24(1).