Transformation of E. coli

Dower, W. J., Miller, J. F. and Ragsdale, C. W. (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 16, 6127-6145. 

HIV-1 integrase can be purified by transformation of E. coli strain BL21... 

To accomplish the many production schemes described in this chapter, genetic transformation of E. coli is necessary. Genetic transformation falls broadly into two categories, either increase of expression for a given polypeptide (overexpression) or decrease (knockdown or knockout), which are used to direct metabolite flow for metabolic engineering purposes. There are many effective strategies for each type of transformation. Bacterial transformation has been covered extensively elsewhere [205], so it is not covered at length here. 

Transformation of E. coli, Production of Much Monomer Gene and Verification of Monomer Gene Sequence... 

Leave the ligation mixture at room temperature for 5 min, and then use it directly for transformation of E. coli competent cells, or store it by freezing until transformation. 

We have not been able to detect any accumulation of medium chain fatty acids in 35S-MCTE transformed Brassica leaves, even though leaf MCTE activity in vitro was comparable to OTE activity. There are two possible explanations for this either the medium chain fatty acids do not accumulate because they are metabolised immediately, most likely via P -oxidation. MCTE transformation of E. coli only results in accumulation of medium chain fatty acids in mutants of E.coli that cannot P-oxidise. Alternatively, in vitro measurement of MCTE activity may not be a true reflection of leaf MCTE activity in vivo. In the chloroplast acyl-ACP substrates may not be accessible to MCTE, a protein that is usually associated with plastid fatty acid synthesis. 

With time, the microbial chip fabrication procedures have evolved enabling SECM studies of increasingly complex biological processes. Nagamine et al. reported an on-chip transformation of E. coli cells. Their technique was based on soft lithographic processes, which were... 

Kaiser, A. D., Hogness, D. The transformation of E. coli with DNA isolated from bacteriophage Mg, J. molec. Biol. 2, 392-415 (i960). 

---