Construction of new cloning vehicles with genes of the tryptophan operon of Escherichia coli as genetic markers
In vitro recombination techniques were used to construct a series of new cloning vehicles with genes of the tryptophan ( trp) operon of Escherichia coli as selective marker. To construct these plasmids we have made a restriction cleavage map of the trp operon for the enzymes AosI, AvaI, BglI, BglII,...
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Published in | Gene Vol. 9; no. 1; pp. 69 - 85 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.04.1980
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Subjects | |
Online Access | Get full text |
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Summary: | In vitro recombination techniques were used to construct a series of new cloning vehicles with genes of the tryptophan (
trp) operon of
Escherichia coli as selective marker. To construct these plasmids we have made a restriction cleavage map of the
trp operon for the enzymes
AosI,
AvaI,
BglI,
BglII,
HindIII,
HpaI,
PvuII,
SalI,
SstI and
XhoI.
The constructed plasmids pHP39, pEP392, pEP3921 and pEP3923 are derived from the amplifiable plasmid pBR345 and carry two or more genes of the
trp operon, which are controlled by the
trp regulatory elements. Plasmid pEP3921 (7.0 kb) carries intact
trpE and
trpA genes and contains single
BglII and
SstI sites in
trpE, a single
HindIII site located between
trpE and
trpA, and single
EcoRI,
SalI and
XhoI sites located outside the
trp genes. Plasmid pEP121 (12 kb) is similar to pEP3921, but has an extra selective marker conferring bacterial resistance to ampicillin. Plasmid pEP3923 (7.4 kb) comprises intact
trpB and
trpA genes and single
BglII,
HindIII,
EcoRI,
SalI and
XhoI sites. Plasmids pHP39 (9.8 kb) and pEP392 (9.8 kb) carry an intact
trp operon and have two and one
EcoRI site, respectively. Plasmid pHP3 (18 kb) carries an intact
trp operon and markers for tetracycline and ampicillin resistance. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0378-1119 1879-0038 |
DOI: | 10.1016/0378-1119(80)90167-5 |