| Chapter title |
The Expression, Purification, and Structure Determination of BamA from E. coli
|
|---|---|
| Chapter number | 13 |
| Book title |
The BAM Complex
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2871-2_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2870-5, 978-1-4939-2871-2
|
| Authors |
Dongchun Ni, Yihua Huang, Ni, Dongchun, Huang, Yihua |
| Abstract |
In gram-negative bacteria, assembly of outer membrane proteins requires the multicomponent β-barrel assembly machinery (BAM) complex, of which BamA is an essential and evolutionarily conserved integral outer membrane protein. To understand how BamA facilitates outer membrane protein biogenesis, it is important to obtain sufficient amounts of purified recombinant BamA protein for in vitro functional analysis and structure determination. In this chapter, we describe the protocol that we used in our laboratory for the cloning, expression, and purification of E. coli BamA and its N-terminal deletion variants for in vitro functional studies and for structure determination of the β-barrel domain alone (residues 426-810). |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 13 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 31% |
| Student > Bachelor | 2 | 15% |
| Professor | 1 | 8% |
| Researcher | 1 | 8% |
| Unknown | 5 | 38% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 31% |
| Arts and Humanities | 1 | 8% |
| Agricultural and Biological Sciences | 1 | 8% |
| Immunology and Microbiology | 1 | 8% |
| Medicine and Dentistry | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 4 | 31% |