Chromosome Abnormalities

1
Chromosome Abnormalities
3.1
2

• Candidate genes pre-genome
• Use eg cosmid fragments to look for genes.
• Must hybridize to RNA on Northerns or in cDNA
  libraries
• CpG islands. Cluster of rare-cutting restriction
  sites implies CpG rich region at promoter of
  gene
• Zoo blots genes are likely to be conserved, so
  use probes on different species
• Specific transcripts in expected tissues, eg
  DMD, look in muscle.
• Isolate cDNA, look for ORF and mutation in
  patients

3.2
3
Dystrophin
CFTR
From Human Molecular Genetics (P Sudbery)
3.3
4
CFTR
5
Comparison of pre and post genome gene hunting
3.4
6
Cloning of the Artemis gene
1. Linkage analysis localises to 10p close to 2
markers
2. Two BAC contigs in this region
3. Sequence prediction identified a candidate
peptide
4. EST blast identified EST contig 1
5. Sequence.
6. Blast with new sequence identified EST contig 2
7. Predicted cDNA sequence checked with indicated
PCR primers
3.5