| Chapter title |
The BAM Complex
|
|---|---|
| Chapter number | 18 |
| Book title |
The BAM Complex
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2871-2_18 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2870-5, 978-1-4939-2871-2
|
| Authors |
Ricci, Dante P, Dante P. Ricci, Ricci, Dante P. |
| Abstract |
The central Bam components BamA and BamD are both essential genes in E. coli, a fact that often confounds genetic analysis using classical methods. The isolation of "depletion strains" in which these genes can be conditionally expressed removes this obstacle and facilitates the in vivo characterization of Bam function. This chapter describes an efficient two-step recombineering method for the construction of such a depletion strain, which contains an arabinose-inducible allele of bamD, using the λ Red system. Additionally, a simple protocol is presented for the depletion of bamD expression in live cells, which is particularly useful for the characterization of mutant alleles of bamD (complementation analysis). In principle, the procedures described can be adapted to produce and characterize depletion strains for any essential gene in E. coli or any other bacterium that is similarly amenable to genome engineering. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 23 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 8 | 35% |
| Student > Doctoral Student | 2 | 9% |
| Student > Bachelor | 2 | 9% |
| Researcher | 2 | 9% |
| Student > Master | 2 | 9% |
| Other | 1 | 4% |
| Unknown | 6 | 26% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 10 | 43% |
| Agricultural and Biological Sciences | 2 | 9% |
| Environmental Science | 1 | 4% |
| Immunology and Microbiology | 1 | 4% |
| Earth and Planetary Sciences | 1 | 4% |
| Other | 1 | 4% |
| Unknown | 7 | 30% |