| Chapter title |
Hexahistidine-tagged maltose-binding protein as a fusion partner for the production of soluble recombinant proteins in Escherichia coli.
|
|---|---|
| Chapter number | 11 |
| Book title |
High Throughput Protein Expression and Purification
|
| Published in |
Methods in molecular biology, January 2009
|
| DOI | 10.1007/978-1-59745-196-3_11 |
| Pubmed ID | |
| Book ISBNs |
978-1-58829-879-9, 978-1-59745-196-3
|
| Authors |
Austin, Brian P, Nallamsetty, Sreedevi, Waugh, David S, Brian P. Austin, Sreedevi Nallamsetty, David S. Waugh |
| Abstract |
Insolubility of recombinant proteins in Escherichia coli is a major impediment to their production for structural and functional studies. One way to circumvent this problem is to fuse an aggregation-prone protein to a highly soluble partner. E. coli maltose-binding protein (MBP) has emerged as one of the most effective solubilizing agents. In this chapter, we describe how to construct combinatorially-tagged His(6)MBP fusion proteins by recombinational cloning and how to evaluate their yield and solubility. We also describe a procedure to determine how efficiently a His(6)MBP fusion protein is cleaved by tobacco etch virus (TEV) protease in E. coli and a method to assess the solubility of the target protein after it has been separated from His(6)MBP. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 4 | 8% |
| Austria | 1 | 2% |
| Unknown | 45 | 90% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 11 | 22% |
| Student > Ph. D. Student | 10 | 20% |
| Student > Bachelor | 8 | 16% |
| Student > Master | 6 | 12% |
| Student > Doctoral Student | 3 | 6% |
| Other | 5 | 10% |
| Unknown | 7 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 17 | 34% |
| Biochemistry, Genetics and Molecular Biology | 15 | 30% |
| Chemistry | 5 | 10% |
| Mathematics | 1 | 2% |
| Physics and Astronomy | 1 | 2% |
| Other | 3 | 6% |
| Unknown | 8 | 16% |