Please use this identifier to cite or link to this item: http://hdl.handle.net/1946/8649
Cloning of TCEA1, TCEA1Mut and TCEA1Del genes into mammalian expression vector pcDNA4/TO
The RNA polymerase II complex transcribes all protein coding genes in the eukaryotic genome. Its functionality depends on multiple transcription factors and subunits, including the elongation factor TFIIS. TFIIS stimulates recovery of backtracked RNA polymerase complexes by transcript cleavage, and loss of function mutations can cause transcriptional blocking of genes with backtracked polymerases. In S. Cerevisiae this elongation factor is not neseccary for cell survival but without it polymerase recovery is a much slower process and this can effect cell viability. Interestingly mutations in certain domains of the yeast TFIIS can not only stop its catalytic effect on transcript cleavage but also inhibit unstimulated RNA polymerase recovery, thus having a leathal effect. To study the effect of mutations in the human TFIIS, encoded by the TCEA1 gene on chromosome 8, the gene has been mutated and has to be cloned into a mammalian expression vector.