A novel strategy to identify the location of necessary and sufficient cis-acting regulatory mRNA elements in trypanosomes.

ABSTRACT: Expression of nearly all protein coding genes in trypanosomes is regulated post-transcriptionally, predominantly at the level of mRNA half-life. The identification of cis-acting elements involved in mRNA stability has been hindered by a lack of ability to screen for loss-of-regulation mutants. The method described in this article allows the region containing the necessary and sufficient elements within a mRNA to be identified and uses antibiotic resistance genes as both selectable markers and reporters. In the case of unstable mRNAs, the strategy can be extended by performing a screen for spontaneous loss-of-function mutants in regulatory parts of a mRNA. The method was validated by using the GPI-PLC mRNA, which is unstable in procyclic form trypanosomes and showed that the 3'UTR of the GP

SUBMITTER: Webb H 

PROVIDER: S-EPMC1360220 | biostudies-literature | 2005 Jul

REPOSITORIES: biostudies-literature