Expression and Purification of a Novel Anti-Tumor Recombinant Fusion Protein Consisting of Trichosan

来源 :BITs 3rd Annual World Cancer Congress-2012(2012第五届世界癌症大会) | 被引量 : 0次 | 上传用户:lzzhong9910
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
  Trichosanthin (TCS), originally extracted from a herbs of traditional Chinese medicine, is a type Ⅰ ribosome-inactivating protein.It has been well-established that TCS has evident anti-tumor effects largely by inducing apoptosis of cancer cells and currently TCS is an approved clinical drug in China.However, the clinical application of native TCS is limited by its poor tumor-targeting and cell-penetrating due to the various distribution of its specific cellular receptors.To address this issue, a genetically engineered TCS was constructed in which TCS was fused with a recently invented supercharged green fluorescent protein (scGFP, provided by Dr.David Liu of Harvard University) based on the new discovery that scGFP-tagged fusion protein acquired ability to penetrate cells.The fusion gene of TCS-scGFP was inserted into the prokaryotic expression vector pET (Novagen) and the recombinant plasmid was transformed into E.coli BL21 (DE3)pLysS, then the fusion protein expression was induce by IPTG and subsequently purified by Ni-NTA system following Qiagens protocol.It turned out that a novel fusion protein TCS-scGFP was prokaryotically expressed and purified successfully, as demonstrated by Western blot.In addition, the experiments of Coomassie brilliant blue staining and density scanning showed that the expressed protein accounted for >50% of total proteins.The pilot experiment indicated that the TCS-scGFP fusion protein was able to transduce HeLa cancer cells efficiently.It is expected the novel TCS-scGFP fusion protein will overcome the limitations of native TCS and the further studies are in progress.
其他文献
Lung cancer remains the leading cause of cancer death in the United States and worldwide.More than 200,000 Americans died every year of lung cancer.At the time of presentation of Non-Small Cell Lung C
会议
The successful development of the YICHAO Meridian is based on a Meridian theory of traditional Chinese medicine,and referred to a variety of manufacturing principles of modem medical devices, which we
会议
Hypofractioned radiation therapy regiments are increasing being used to treat patients desiring breast conserving therapy.Shortened treatment regiments have been shown to improve quality of life and i
会议
Objectives: In this study, we used female rats, which were treated with 7,12-dimethylbenz(a)anthracene (DMBA) to induce breast tumors and with the concomitant of alcohol intake.The role of alcohol in
会议
In this study, perioperative outcomes and survival data in patients with cervical cancer operated with three surgical methods: robot-assisted, laparoscopic and open, are to be analyzed.From January 20
会议
For many decades, adjuvant radiotherapy has been used after mastectomy for patients with locally advanced breast cancer.By reducing the risk of recurrency, radiotherapy can improve the overall surviva
会议
Chemotherapy-induced nausea and vomiting (CINV) is experienced by around 80% of patients currently receiving chemotherapy.Acute CINV typically occurs within minutes or hours of drug administration, la
会议
Background: Multidrug resistance is the most frequent type of resistance to anticancer chemotherapy and diseasemodifying antirheumatic drugs (DMARDs), which usually results from the overexpression of
会议
Purpose:The aim of this study was to evaluate i) differences in the HLA groups of renal cell carcinoma (RCC) patients and healthy control persons and ii) to study association of expression of uPA-fami
会议
The present study was conducted to verify whether caffeine is beneficial for improving leukaemia therapy.Co-treatment with adaphostin (a Bcr/Abl inhibitor) was found to potentiate caffeine induced Fas