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It was originally thought that Escherichia coli, a commonly used laboratory organism, was refractory to transformation. However, in 1970, Morton Mandel and Akiko Higa showed that E. coli may be induced to take up DNA from bacteriophage λ without the use of helper phage after treatment with calcium chloride solution. [6]
Evolution in bacteria was previously viewed as a result of mutation or genetic drift. [11] Today, genetic exchange, or gene transfer is viewed as a major driving force in the evolution of prokaryotes. [11] This driving force has been widely studied in organisms like E. coli. [12]
Genetic systems have also been developed which allow the production of recombinant proteins using E. coli. One of the first useful applications of recombinant DNA technology was the manipulation of E. coli to produce human insulin. [26] Modified E. coli have been used in vaccine development, bioremediation, and production of immobilised enzymes ...
E. coli BL21(DE3) lacks a functional type I restriction-modification system, indicated by hsdS(r B-m B-). Specifically, both the restriction (hsdR) and modification (hsdM) domains are inactive. This enhances transformation efficiency since exogenously introduced unmethylated DNA remains untargeted by the restriction-modification system. [9]
During classical E. coli conjugation the donor cell provides a conjugative or mobilizable genetic element that is most often a plasmid or transposon. [5] Most conjugative plasmids have systems ensuring that the recipient cell does not already contain a similar element. The genetic information transferred is often beneficial to the recipient.
Calcium chloride treatment is generally used for the transformation of E. coli and other bacteria. [11] It enhances plasmid DNA incorporation by the bacterial cell, promoting genetic transformation. Plasmid DNA can attach to LPS by being added to the cell solution together with CaCl 2. [12]
Microbial genetics is a subject ... Gene transfer systems that have been extensively studied in bacteria include genetic transformation, ... E. coli conjugation ...
A bacterial artificial chromosome (BAC) is a DNA construct, based on a functional fertility plasmid (or F-plasmid), used for transforming and cloning in bacteria, usually E. coli. [1] [2] [3] F-plasmids play a crucial role because they contain partition genes that promote the even distribution of plasmids after bacterial cell division.