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| 人羧肽酶A1的毕赤酵母可溶表达 | |
| 引用本文： | 易静,刘家云,缪应叶,张青,苏明权,岳乔红,郝晓柯.人羧肽酶A1的毕赤酵母可溶表达[J].西安交通大学学报(医学版),2006,27(4):324-326,336. |
| 作者姓名： | 易静 刘家云 缪应叶 张青 苏明权 岳乔红 郝晓柯 |
| 作者单位： | 1. 第四军医大学西京医院检验科,陕西,西安,710033 2. 解放军152医院检验科,河南,平顶山,467000 3. 第四军医大学唐都医院检验科,陕西,西安,710038 |
| 基金项目： | 国家“863”高技术研究发展计划基金资助项目(No.2001AA215321) |
| 摘 要： | 目的应用毕赤酵母表达系统表达人羧肽酶A1(carboxypeptidase A1,CPA1)蛋白。方法从pGEX-CPA1重组质粒中扩增出CPA1基因,亚克隆入酵母表达载体pPIC9K,酵母重组表达载体pPIC9K-CPA1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-CPA1,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蛋白。结论在毕赤酵母中成功表达了CPA1,为进一步研究CPA1在抗体导向酶-前体药物疗法中的应用奠定了基础。 |
| 关 键 词： | 羧肽酶A1 巴斯德毕赤酵母 基因表达 |
| 文章编号： | 1671-8259（2006）04-0324-03 |
| 收稿时间： | 2006-02-24 |
| 修稿时间： | 2006-02-242006-03-16 |
Soluble expression of human carboxypeptidase A1 active center in pichia pastoris |
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| Yi Jing,Liu Jiayun,Miao Yingye,Zhang Qing,Su Mingquan,Yue Qiaohong,Hao Xiaoke.Soluble expression of human carboxypeptidase A1 active center in pichia pastoris[J].Journal of Xi‘an Jiaotong University:Medical Sciences,2006,27(4):324-326,336. | |
| Authors: | Yi Jing Liu Jiayun Miao Yingye Zhang Qing Su Mingquan Yue Qiaohong Hao Xiaoke |
| Abstract: | (Objective)To express the human carboxypeptidase A1 in soluble form in pichia pastoris.(Methods)Human carboxypeptidase A1 gene was amplified from pGEX-CPA1 recombinant plasmid and subcloned into pPIC9K yeast expression vector.Recombinant vector pPIC9K-CPA1 was transformed into pichia SMD1168 by electroporation.Positive recombinant yeast strains were screened through nutrition defect and G418 resistance.Expression of the integrated multicopy recombinant yeast strains were induced.Results The yeast recombinant expression vector pPIC9K-CPA1 was constructed.Recombinant yeast strains which integrated over 16 recombinant vector copies and soluble human carboxypeptidase A1 protein were obtained.(Conclusion)The human carboxypeptidase A1 can be expressed successfully in pichia pastoris,which will be useful for further research on ADEPT of prospate. |
| Keywords: | carboxypeptidase A1 pichia pastoris gene expression |
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