9.19.2007

Gene Cloning Methods - Cloning in E. coli - Cloning Bacterial Genes

Gene Cloning Methods - Cloning in E. coli - Cloning Bacterial Genes: "

Top 5 Reasons E. coli is used for Gene Cloning

The microorganism Escherichia coli has a long history of use in the biotechnology industry and is still the microorganism of choice for most gene cloning experiments. Although E. coli is known to the general population for the infectious nature of one particular strain (0157:H7) few people are aware of how versatile and useful E. coli is to genetic research. There are several reasons E. coli became so widely used and is still a common host for recombinant DNA.

1. Genetic Simplicity

Bacteria make useful tools for genetic research because of their relatively small genome size compared to eukaryotes. E. coli cells only have about 4,400 genes whereas humans contain approximately 100,000. Also, bacteria, including E. coli, live their entire lifetime in a haploid state, with no second allele to mask the effects of mutations during protein engineering experiments.

source:

Weaver, R. and Hedrick, P. 1989. Genetics. Wm. C. Brown Publishers, Dubuque, IA, USA.

Madigan, M., Martinko, J. and Parker, J. 2000. Brock Biology of Micro-organisms, 9th ed. Prentice Hall, Upper Saddle River, NJ, USA.

2. Growth Rate

Bacteria typically grow much faster than more complex organisms. E. coli grows rapidly at a rate of one generation per twenty minutes under typical growth conditions. This allows for preparation of log-phase cultures overnight and genetic experimental results in mere hours instead of several days, months or years.

3. Safety

E. coli is naturally found in the intestinal tracts of humans and animals where it helps provide nutrients (vitamins K and B12) to its host. There are many different strains of E. coli that may produce toxins or cause varying levels of infection if injested or allowed to invade other parts of the body. Despite the bad reputation one particularly toxic strain (O157:H7), E. coli are generally relatively inocuous if handled with reasonable hygiene.

4. Conjugation and the Genome Sequence

The E. coli genome was the first to be completely sequenced. Genetic mapping in E. coli was made possible by the discovery of conjugation. E. coli is the most highly studied microorganism and an advanced knowledge of its protein expression mechanisms make it simpler to utilize for experiments where expression of foreign proteins and selection of recombinants is essential.

source:

Weaver, R. and Hedrick, P. 1989. Genetics. Wm. C. Brown Publishers, Dubuque, IA, USA.

5. Ability to Host Foreign DNA

Most gene cloning techniques were developed using this bacterium and are still more successful or effective in E. coli than in other microorganisms. E. coli is readily transformed with plasmids and other vectors, and preparation of competent cells is not complicated. Transformations with other microorganisms are often less successful.

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