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. 1975 Nov;72(11):4361–4365. doi: 10.1073/pnas.72.11.4361

Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome.

L Clarke, J Carbon
PMCID: PMC388721  PMID: 1105581

Abstract

Using a poly(dA-dT) "connector" method, a population of annealed hybrid circular DNAs was constructed in vitro; each hybrid DNA circle containing one full-length molecule of poly(dT)-tailed DNA from E1 colicinogenic factor (Col E1) fragmented by EcoRI endonuclease annealed to any one of a collection of poly(dA)-tailed linear DNA fragments of the entire E. coli genome. This annealed, but unligated, hybrid DNA was used to transform several different auxotrophic mutants of E. coli, and by direct selection, bacterial clones were isolated which contained specific hybrid plasmids. In this manner, bacterial strains containing Col E1 hybrid plasmids carrying the entire tryptophan operon or the arabinsoe and leucine operons were isolated. The methods described should allow the molecular cloning of any portion of the E. coli genome by selection from a pool of DNA molecules containing at least several hundred different hybrids representing the entire bacterial genome.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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