Knowledge Synthesize System Base on Research Document
| Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1) | |
| Galluccio, Michele1; Pantanella, Marta1; Giudice, Deborah1; Brescia, Stefania1; Indiveri, Cesare1,2 | |
| 2020-08 | |
| 发表期刊 | MOLECULAR BIOLOGY REPORTS
 |
| ISSN | 0301-4851 |
| EISSN | 1573-4978 |
| 摘要 | It is well established thatEscherichia colirepresents a powerful tool for the over-expression of human proteins for structure/function studies. In many cases, such as for membrane transporters, the bacterial toxicity or the aggregation of the target protein hamper the expression limiting the application of this tool. The aim of this study was finding the appropriate conditions for the expression of reluctant proteins that is the human neutral amino acid transporters ASCT2 and B0AT1, that have great relevance to human health in cancer therapy and in COVID-19 research, respectively. The cDNAs coding for the proteins of interest were cloned in the pCOLD I vector and differentE. colistrains (BL21 codon plus RIL, and RosettaGami2) were cultured in absence or in presence of glucose (0.5-1%), at low temperature (15 degrees C), and low inducer concentrations (10-100 mu M). Cell growth and protein production were monitored by optical density measurements and western blotting assay, respectively. Even though in different conditions, the expression of both amino acid transporters was obtained.Reducing the growth rate of specificE. colistrains by lowering the temperature and the IPTG concentration, together with the addition of glucose, two reluctant human neutral amino acid transporters have been expressed inE. coli. The results have a potentially great interest in drug discovery since ASCT2 is an acknowledged target of anticancer therapy, and B0AT1 together with ACE2 is part of a receptor for the SARS-Cov-2 RBD proteins. |
| 关键词 | Amino acid SLC Protein expression B0AT1 ASCT2 COVID-19 |
| DOI | 10.1007/s11033-020-05717-8 |
| WOS关键词 | ESCHERICHIA-COLI ; MEMBRANE TRANSPORTERS ; OVER-EXPRESSION ; B(0)AT1 SLC6A19 ; STRATEGIES ; PROTEINS ; HEALTH |
| WOS研究方向 | Biochemistry & Molecular Biology |
| WOS类目 | Biochemistry & Molecular Biology |
| 出版者 | SPRINGER |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 专题 | 新冠肺炎 循证社会科学证据集成 |
| 作者单位 | 1.Univ Calabria; 2.CNR Inst Biomembranes Bioenerget & Mol Biotechnol |
| 推荐引用方式 GB/T 7714 | Galluccio, Michele,Pantanella, Marta,Giudice, Deborah,et al. Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1)[J]. MOLECULAR BIOLOGY REPORTS,2020. |
| APA | Galluccio, Michele,Pantanella, Marta,Giudice, Deborah,Brescia, Stefania,&Indiveri, Cesare.(2020).Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1).MOLECULAR BIOLOGY REPORTS. |
| MLA | Galluccio, Michele,et al."Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1)".MOLECULAR BIOLOGY REPORTS (2020). |
| 条目包含的文件 | 下载所有文件 | |||||
| 文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
| Galluccio-2020-Low t（918KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | 浏览 下载 | |
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。
修改评论