Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Molecular Plant Breeding》 2018-10
Add to Favorite Get Latest Update

Molecular Cloning of Cassava MeCREB Gene and Its Expression and Condition Optimization in Prokaryotic Cells

Wang Shuo;Liu Jiao;Fu Shaoping;Duan Ruijun;Li Ruimei;Yao Yuan;Hu Xinwen;Guo Jianchun;Institute of Tropical Agriculture and Forestry, Hainan University;Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture,Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;  
MeCREB gene was cloned by RT-PCR from cassava in this study. The analysis of nucleic acid sequence indicated that the cloned gene was consistent with the sequence of MeCREB gene(Manes. 04 g058300.1)recorded in the phytozome database. The MeCREB gene was 968 bp with an open reading frame(ORF) of 747 bp which encoded 248 amino acids. MeCREB protein was predicted to be an unstable hydrophilic protein, encoding a protein of 26.26 kD. The theoretical isoelectric point of MeCREB was 5.99, and the GRAVY was-0.667. The MeCREB-pGEX-6 p-1 expression vector was constructed and induced to express in E. coli. Also, the expression conditions such as temperature, bacterium concentration, IPTG concentration and induction time were optimized by Me C REB-p GEX-6 p-1 to confirm the optimal conditions for the expression of GST-MeCREB fusion protein.The results showed that the conditions of 28℃, OD600=0.8, 0.1 mmol/L concentration of IPTG and 6 hours' culture were the most suitable for the expression of GST-MeCREB fusion protein. This study provided an optimal scheme for efficiently getting the GST-MeCREB fusion protein using for the downstream experiments, and laid the foundation for the further study on the biological functions of MeCREB genes.
【Fund】: 国家自然科学基金(No.31371706);; 中国科协青年人才托举工程项目(No.YESS20150167)共同资助
【CateGory Index】: Q943.2;S533
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved