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| 大鼠血红素加氧酶-1基因在乳酸乳球菌中的克隆与表达 | |
| 引用本文： | 吴长毅,曾因明,王俊科,庞庆丰,杨光,高新跃. 大鼠血红素加氧酶-1基因在乳酸乳球菌中的克隆与表达[J]. 应用与环境生物学报, 2006, 12(1): 48-51 |
| 作者姓名： | 吴长毅 曾因明 王俊科 庞庆丰 杨光 高新跃 |
| 作者单位： | 1. 中国医科大学第一临床学院,沈阳,110001;江苏省麻醉医学研究所基因工程研究室,江苏,徐州,221002 2. 江苏省麻醉医学研究所基因工程研究室,江苏,徐州,221002 3. 中国医科大学第一临床学院,沈阳,110001 |
| 基金项目： | Acknowledgements We thank Prof. Luis （ Unite de Recherches Laitie＇res et de Ge ne tique Applique e, France） for providing plasmid pSEC-E7 and L. lactis strain NZ9000. |
| 摘 要： | 采用RTPCR技术从大鼠脾总RNA中分离扩增HO1基因,将该基因克隆进pGEMTeasy质粒中,转化大肠杆菌DH5α,提取质粒,分析基因,酶切后与含有乳酸乳球菌启动子NisA的pSEC质粒连接,经电击转化,将重组质粒转入乳酸乳球菌NZ9000中,转化子在含有氯霉素的脑心浸液培养基上培养.用Nisin诱导HO1表达,SDSPAGE和Westernblot分析、鉴定表达产物,并且用分光光度法测定工程菌表达的HO1活性为0.45nmolmg(protein)-1h-1.图5参18 |
| 关 键 词： | 乳酸乳球菌 血红素加氧酶-1 基因克隆表达 |
| 收稿时间： | 2005-06-02 |
| 修稿时间： | 2005-09-20 |
Cloning and Expression of Rat Heme Oxygenase-1 Gene in Lactococcus lactis |
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| WU Changyi,ZENG Yinming,WANG Junke,PANG Qingfeng,YANG Guang,GAO Xinyue. Cloning and Expression of Rat Heme Oxygenase-1 Gene in Lactococcus lactis[J]. Chinese Journal of Applied and Environmental Biology, 2006, 12(1): 48-51 | |
| Authors: | WU Changyi ZENG Yinming WANG Junke PANG Qingfeng YANG Guang GAO Xinyue |
| Affiliation: | 1.Department of Anesthesiology, First Affiliated Hospital, China Medical University, Shenyang 110001, China; 2 .Department of Gene Engineering, Jiangsu Institute of Anesthesiology, X uzhou 221002, Jiangsu, China |
| Abstract: | Total RNA was extracted from rat spleen tissue using RNA kit and heme oxygenase-1 ( HO-1 ) gene was amplified by RT-PCR. The sequence of HO-1 gene was identified by T/A clone and DNA sequencing. Then, cDNA of HO-1 was inserted into the NisⅠ/EcoRⅠ site of the pSEC vector with PnisA promoter, resulting in the pSEC-HO-1 expressing vector. The vector was transformed into Lactococcus lactis NZ9000 by electroporation, and the expression of HO-1 gene induced by nisin was evaluated by SDS-PAGE and Western blot. In addition, the activity of HO-1 expressed by engineering L. lactis was 0.45 nmol mg (protein) - 1 h - 1 using spectrophotography. Fig 5, Ref 18 |
| Keywords: | Lactococcus lactis heme oxygenase-1 gene cloning and expression |
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