The following information accompanied stocks donated to the Bloomington Stock Center by Kent Golic, The University of Utah.
The generation of the p53 allele p535A-1-4 is described in Xie and Golic, 2004, Genetics 168(3): 1477--1489 (FBrf0180287).
The generation of the p53 allele p5311-1B-1 is described in Rong et al., 2002, Genes Dev. 16(12): 1568--1581 (FBrf0149003).
The two p53- alleles, p5311-1B-1 and p535A-1-4 have no obvious phenotype.
The structure of the alleles can be verified by PCR.
p535A-1-4 is a deletion. These primers:
P53-C1: 5'-AGCTAATGTGACTTCGCATTGAACAAA-3'
P53-C2: 5'-TCGATAAACATTGGCTACGGCGATTGT-3'
will give a 4kb product from the deletion and a 7.3kb product from the wild-type allele.
p5311-1B-1 results from introduction of a stop mutation into the coding sequence. It can be verified by allele-specific PCR with the following three primers:
(1) 5'-GTTCGCCTGGATCTTAATTA-3';
(2) 5'-GTTCGCCTGGATCTGAATGT-3'; and
(3) 5'-AATCGCTGCATGCGGTAGTA-3'.
Primers 1 and 3 generate a 1.3kb fragment specifically from the mutant allele.
Primers 2 and 3 generate a 1.3kb fragment specifically from the wild-type allele.