| 根癌农杆菌介导脂肪酶在无孢黑曲霉中的高效表达 |
| |
| 引用本文： | 潘力,王云艳,王斌,何攀,李俊星.根癌农杆菌介导脂肪酶在无孢黑曲霉中的高效表达[J].华南理工大学学报(自然科学版),2012,40(5):84-89. |
| |
| 作者姓名： | 潘力 王云艳 王斌 何攀 李俊星 |
| |
| 作者单位： | 华南理工大学生物科学与工程学院,广东广州,510006 |
| |
| 基金项目： | 国家自然科学基金资助项目 |
| |
| 摘 要： | 黑曲霉(Aspergillus niger)具有高效的蛋白表达和分泌能力.为实现异源蛋白在无孢黑曲霉中的高效重组表达,在优化遗传筛选标记的基础上,建立了根癌农杆菌介导的、以潮霉素抗性基因为筛选标记的黑曲霉转化体系.利用该体系介导米黑根毛霉脂肪酶(RML)转化黑曲霉SH-1,通过PCR鉴定、酶活检测、镍柱亲和层析及SDS-PAGE电泳检测等确定成功获得了RML的黑曲霉转化子,并通过发酵条件优化实现了RML的高效表达,对硝基苯酚比色法测得转化子酶活最高可达15 U/mL,是其在酵母中表达水平的10倍以上.
|
| 关 键 词： | 无孢黑曲霉 根癌农杆菌介导转化 米黑根毛霉脂肪酶 基因表达 |
Effective Expression of Lipase in Non-Spore Aspergillus niger via Agrobacterium tumefaciens-Mediated Transformation |
| |
| Pan Li
,
Wang Yun-yan
,
Wang Bin
,
He Pan
,
Li Jun-xing.Effective Expression of Lipase in Non-Spore Aspergillus niger via Agrobacterium tumefaciens-Mediated Transformation[J].Journal of South China University of Technology(Natural Science Edition),2012,40(5):84-89. |
| |
| Authors: | Pan Li Wang Yun-yan Wang Bin He Pan Li Jun-xing |
| |
| Institution: | Pan Li Wang Yun-yan Wang Bin He Pan Li Jun-xing(School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,Guangdong,China) |
| |
| Abstract: | Aspergillus niger(A.niger) owns effective capabilities of protein expression and secretion.In order to implement the effective recombinant expression of heterologous protein in non-spore A.niger,several selection mar-kers were evaluated and hygromycin B resistance gene was selected to establish an Agrobacterium tumefaciens(A.tumefaciens)-mediated transformation system of A.niger.Then,Rhizomucor miehei lipase(RML) was transformed into A.niger SH-1 via the A.tumefaciens-mediated transformation and was verified by means of PCR identification,enzyme activity assay,Ni affinity chromatography and SDS-PAGE.Finally,RML transformants effectively expres-sing RML protein were obtained after the fermentation optimization.The p-nitrophenol colorimetry results show that the transformants possesses a lipase activity of 15 U/mL,which is more than ten times that expressed in yeast. |
| |
| Keywords: | non-spore Aspergillus niger Agrobacterium tumefaciens-mediated transformation Rhizomucor miehei lipase gene expression |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
