HELP

MICCOLLERAN at cs8700.ucg.ie MICCOLLERAN at cs8700.ucg.ie
Fri Jun 28 21:09:00 EST 1991


I wish this letter to be distributed around the world to Bionews notice boards.

To anyone with experience in the cloning of beta-glucosidase genes from
eubacteria I would like to ask you for your help.

We are currently engaged in a project to isolate a gene encoding a linamarase
enzyme from Leuconostoc mesenteroides.   We have constructed a representative
genomic library of L. mesenteroides in the Lambda phage EMBL4.   We require an
assay to isolate clones bearing beta-glucosidases and more particularly
linamarases.   We would be grateful to anyone who would have a partial or total
   sequence, be it protein sequence or other, of a linamarase from any organism.
      We wish simply to generate a nucleic acid probe to the linamarase locus in

L.mesenteroides.   We will respect the confidentiallity of all replies.

Linamarin is a beta-glucose with a cyanide group attached.   We have found that
   there is extremely little nucleotide sequence conservation within
beta-glucosidases.   We cannot generate a probe from beta-glucosidases.

An alternative approach to the study would be the use of
(1) a chemical assay for linamarase containing lambda phage.   Does anyone
in the community know of one please.
(2) screening the genomic library by subtractive hybridization using a linamarin
    negative L. mesenteroides genome as the probe.

We would honestly appreciate any help, advice, sequences and again respect
the confidentiality of all replies.

Yours sincerely,



GERARD J. COLLERAN,
LABORATORY OF PROFESSOR L. K. DUNICAN,
DEPATRMENT OF MICROBIOLOGY,
UNIVERSITY COLLEGE,
GALWAY,
IRELAND.



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