Isolation of plant transcription factors using a modified yeast one-hybrid system.

ABSTRACT:

Background

The preparation of expressional cDNA libraries for use in the yeast two-hybrid system is quick and efficient when using the dedicated Clontechtrade mark product, the MATCHMAKER Library Construction and Screening Kit 3. This kit employs SMART technology for the amplification of full-length cDNAs, in combination with cloning using homologous recombination.Unfortunately, such cDNA libraries prepared directly in yeast can not be used for the efficient recovery of purified plasmids and thus are incompatible with existing yeast one-hybrid systems, which use yeast transformation for the library screen.

Results

Here we propose an adaptation of the yeast one-hybrid system for identification and cloning of transcription factors using a MATCHMAKER cDNA library. The procedure

SUBMITTER: Lopato S 

PROVIDER: S-EPMC1402289 | biostudies-literature | 2006 Feb

REPOSITORIES: biostudies-literature