Two
her-1 transcripts have been previously identified: a 0.8 kb transcript found at high levels in XO and very low levels in XX animals and a 1.2 kb transcript observed at low levels in XO animals only. The dominant
her-1 mutations
n695 and
y101 ,which cause variable masculinization of XX animals, result in increased levels of both
her-1 transcripts in XX embryos. We have cloned and sequenced the
her-1 genes from partial genomic libraries of these mutant strains. Both strains have a G --> A transition a few base pairs upstream of the cap site for the 1.2 kb transcript. That the two strains have the same mutation was not expected since
y101 shows a more extreme sexual transformation than
n695 .The mutation has been confirmed in both strains by direct sequencing of PCR products made from genomic DNA. We have sequenced the exons for each mutant gene and about l.5kb upstream of the transcription start site, but have not observed an additional mutation in either strain. We are in the process of sequencing the recessive
her-1 alleles that result in the preferential loss of the 1.2kb transcript:
ct50 n695 ,
e1519 ,
e1574 ,
n827 n695 and
y10 .In such strains, the 1.2kb transcript cannot be detected by Northern analysis, but the 0.8kb transcript is present at approximately wild-type levels. The
ct50 n695 mutation, used originally to isolate
her-1 sequences, results from a Tc1 insert 55 bp upstream of the cap site for the 1.2 kb transcript. We are analyzing the other four mutant alleles by direct sequencing of DNA generated by PCR amplification of specific regions of the
her-1 gene. The loss of the larger transcript in
e1519 is due to a splicing defect resulting from a G -->A transition mutation in the first base of intron #1. Work is still in progress on the other three alleles.