Cloning， Expressing of Caenorhabditis elegans p53/cep-1 and the Effect on Lifespan

Abstract: Caenorhabditis elegans cep-1 was similar to its mammalian counterpart， tumor suppressor p53. cep-1 was amplified from C.elegans cDNA by PCR and cloned into expression vector and RNA interference vector. The full length of cep-1 cDNA was 2106bp， containing a 5'untranslated region of 27 bp and a 3'untranslated region of 144 bp. The open reading frame was 1935bp which predicted encoding a 645 amino acids protein. CEP-1 recombinant protein was obtained by IPTG induced in Escherichia coli. At the same time， we investigated the effect of p53/cep-1 on C.elegans lifespan in wild type， daf-2 (e1370)， daf-16 (e1038) strains by RNAi (RNA-mediated interference)， and found that the lifespan of wild type was prolonged significantly in cep-1 RNAi worm， in contrast， the lifespan of the other strains have no difference with the RNAi control worms. And the transcription level of cep- 1 in N2， daf-2 (e1370) and daf-16 (e1038) was determined by RT-PCR.

CSTR: 32200.14.cjcb.2010.03.0018