Mycobacterium tuberculosis RpfA protein expression purification and identification of.doc
- 1、本文档共14页,可阅读全部内容。
- 2、原创力文档(book118)网站文档一经付费(服务费),不意味着购买了该文档的版权,仅供个人/单位学习、研究之用,不得用于商业用途,未经授权,严禁复制、发行、汇编、翻译或者网络传播等,侵权必究。
- 3、本站所有内容均由合作方或网友上传,本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺!文档内容仅供研究参考,付费前请自行鉴别。如您付费,意味着您自己接受本站规则且自行承担风险,本站不退款、不进行额外附加服务;查看《如何避免下载的几个坑》。如果您已付费下载过本站文档,您可以点击 这里二次下载。
- 4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等,请点击“版权申诉”(推荐),也可以打举报电话:400-050-0827(电话支持时间:9:00-18:30)。
查看更多
Mycobacterium tuberculosis RpfA protein expression purification and identification of
PAGE \* MERGEFORMAT 14
Mycobacterium tuberculosis RpfA protein expression purification and identification of
Author: Gao Hui, Bai Yin-lan, Li-Mei Wang, Zhi-Kai Xu, Ying Xue
[Keywords:] RpfA; expression; purification; Mycobacterium tuberculosis
Expression, purification and identification of Mycobacterium tuberculosis RpfA
[Abstract] AIM: To express efficiently Mycobacterium tuberculosis RpfA in E. coli and purify the fusion proteins. METHODS: RpfA gene segments were digested with NdeI and BamHI and cloned into pcDNA3.1 vector. After sequencing, RpfA were subcloned into the prokaryotic expression vector pET19b to get pET19bRpfA. The plasmid pET19bRpfA were transformed into E. coli DE3 and induced by IPTG for its expression. The RpfA fusion protein expression was analyzed by SDSPAGE and confirmed by Western blot. Recombinant (His) 6 fusion proteins were purified via Ni2 NTA affinity chromatography. RESULTS: RpfA gene segments were identical to that GenBank reported (lacking 5 ‘ends of 99 bp). The pET19bRpfA vector expressed RpfA fusion proteins at Mr about 80 ku, which could be caught by anti (His) 6 mAb, antiRv1884 and antiRv2389 immune serum.SDSPAGE analysis showed that the fusion proteins mainly existed in inclusion bodies.The expressed proteins could be purified via Ni2 NTA affinity chromatography in denatured condition. CONCLUSION: The recombinant expression plasmid pET19bRpfA has been constructed and RpfA fusion proteins been successfully expressed in E. coli and purified, which lay a basis for further study of RpfA functions.
[Keywords] RpfA; expression; purification; Mycobacterium tuberculosis
[Abstract] Objective: Expression and purification of fusion proteins RpfA. Methods: RpfA gene fragment containing the plasmid with NdeI and BamH Ⅰ restriction enzyme digestion, and then the target gene fragment was cloned into the vector pcDNA3.1 recombinant vector pcDNA3.1 RpfA, sequencing, and then the target gene fragments were subcloned into the pET19
您可能关注的文档
- Mumps complicated by acute pancreatitis in children with clinical analysis of 93 cases of.doc
- Murine cervical heterotopic heart transplantation a comparison of three surgical.doc
- Murine dendritic cells in vitro and identification of.doc
- Musca domestica sequence-related amplified polymorphism amplification system Establishment and Optimization of.doc
- Musca domestica molecular structure and antimicrobial spectrum of antimicrobial peptide research.doc
- Muscle can absorb non-invasive treatment of rib fractures rib 21 cases of nail.doc
- Murray Dalziel talk about the quality of an outstanding CEO.doc
- Muscle strain and muscle spasms difference between prevention and treatment.doc
- Muscle tissue engineering scaffold to cells and human amniotic epithelial cells of the compatibility.doc
- Muscle relaxants on narcotic prevention of complications.doc
- 中国国家标准 GB 42125.11-2025测量、控制和实验室用电气设备的安全要求 第11部分:用于处理医用材料的灭菌器和清洗消毒器的特殊要求.pdf
- 《GB 42125.11-2025测量、控制和实验室用电气设备的安全要求 第11部分:用于处理医用材料的灭菌器和清洗消毒器的特殊要求》.pdf
- 《GB/T 15237-2025术语工作及术语科学 词汇》.pdf
- GB/T 15237-2025术语工作及术语科学 词汇.pdf
- GB/T 22454-2025企业建模与体系结构 企业建模构件.pdf
- 中国国家标准 GB/T 15237-2025术语工作及术语科学 词汇.pdf
- 中国国家标准 GB/T 22454-2025企业建模与体系结构 企业建模构件.pdf
- 《GB/T 22454-2025企业建模与体系结构 企业建模构件》.pdf
- 中国国家标准 GB/T 18910.64-2025液晶显示器件 第6-4部分:测试方法 带动态背光的液晶显示模块.pdf
- 《GB/T 18910.64-2025液晶显示器件 第6-4部分:测试方法 带动态背光的液晶显示模块》.pdf
文档评论(0)