Vectors encoding alternative antibiotic resistance for use in the yeast two-hybrid system.

Abstract:

:We have altered the antibiotic resistance of the reporter plasmids and the pJG4-5 activation-domain and pEG202 DNA binding-domain plasmids used in the Brent interaction trap/two-hybrid system. These plasmids were each previously ampicillin-resistant, resulting in an inefficient purification of any one plasmid from a yeast strain containing all three plasmids that constitute the complete interaction trap. By creating derivatives of each of these plasmids expressing either kanamycin or chloramphenicol resistance, along with the parent plasmids, we now have the option to use the interaction trap in yeast with three E. coli differentially selectable vectors. This will allow isolation of any one plasmid by purifying all of the interaction trap plasmids from yeast simultaneously and plating E. coli transformed with the plasmids onto the appropriate antibiotic plate to select the particular plasmid of interest.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Watson MA,Buckholz R,Weiner MP

doi

10.2144/96212st02

subject

Has Abstract

pub_date

1996-08-01 00:00:00

pages

255-9

issue

2

eissn

0736-6205

issn

1940-9818

journal_volume

21

pub_type

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