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Cloning of the groE operon of the marine bacterium Vibrio harveyi using a lambda vector

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Kuchanny D., Klein G., Krzewska J., Czyz A., Lipinska B.

Acta Biochimica Polonica

1998

tom 45

nr 1

groES and groEL genes encode two co-operating proteins GroES and GroEL, belonging to a class of chaperone proteins highly conserved during evolution. The GroE chaperones are indispensable for the growth of bacteriophage λ in Escherichia coli cells. In order to clone the groEL and groES genes of the marine bacterium Vibrio harveyi, we constructed the V. harveyi genomic library in the λEMBL1 vector, and selected clones which were able to complement mutations in both groE genes of E. coli for bacteriophage λ growth. Using Southern hybridization, in one of these clones we identified a DNA fragment homologous to the E. coli groE region. Analysis of the nucleotide sequence of this fragment showed that the cloned region contained a sequence in 71.7% homologous to the 3' end of the groEL gene of E. coli. This confirmed that the λ clone indeed carries the groE region of V. harveyi. The positive result of our strategy of cloning with the use of the genomic library in λ vector suggests that the same method might be useful in the isolation of the groE homologues from other bacteria. The V. harveyi cloned groE genes did not suppress thermosensitivity of the E. coli groE mutants.

Ograniczanie wyników

1 Acta Biochimica Polonica

1 Czyz A.

1 Klein G.

1 Krzewska J.

1 Kuchanny D.

1 Lipinska B.

1 1998

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Cloning of the groE operon of the marine bacterium Vibrio harveyi using a lambda vector