The murine lymphomacrophage hybrids ESb, EbF1, EbF2-c4, which express c-fes constitutively, were found by Southern analysis to bear a c-fes deletion of almost 100 bp. The deleted allele was transmitted to the metastatic hybrids by their nonexpressing, poorly metastatic T-lymphoma progenitor Eb, which also has a structurally normal c-fes allele. PCR amplification and sequencing of fes cDNA spanning exons 3-5, where the deletion mapped, ruled out any involvement of coding sequences in the rearrangement. PCR amplification of the as yet unsequenced murine c-fes IVS3 and IVS4 showed they are about 50% longer than their human and feline homologs. Sequencing of IVS4 showed no difference between tumor and control DNA. Sequencing of part of the approximately 2600-bp IVS3 was guided by the restriction analysis of PCR products from control and hybrid DNAs. This showed that differences from the control appeared to be mainly located in the 900-bp HindIII-EcoRI fragment, localized in the middle of IVS3. As all three hybrids had the same restriction map, this fragment was sequenced in one of them (ESb). A run of >200 CA repeats was found in control DNA, and a reduction in the CAn microsatellite accounted for most of the c-fes deletion in the ESb hybrid. Interestingly, the 50% reduction in the size of human and feline c-fes IVS3 as compared with the murine homolog is mostly due to contraction of the same microsatellite.

Microsatellite instability in IVS3 of murine c-fes gene: tumor-associated rearrangement and mammalian divergence / L. Volpi, A. Beghini, D. Rossi, L. Larizza. - In: MAMMALIAN GENOME. - ISSN 0938-8990. - 7:9(1996 Sep), pp. 682-685.

Microsatellite instability in IVS3 of murine c-fes gene: tumor-associated rearrangement and mammalian divergence

L. Volpi
Primo
;
A. Beghini
Secondo
;
L. Larizza
Ultimo
1996

Abstract

The murine lymphomacrophage hybrids ESb, EbF1, EbF2-c4, which express c-fes constitutively, were found by Southern analysis to bear a c-fes deletion of almost 100 bp. The deleted allele was transmitted to the metastatic hybrids by their nonexpressing, poorly metastatic T-lymphoma progenitor Eb, which also has a structurally normal c-fes allele. PCR amplification and sequencing of fes cDNA spanning exons 3-5, where the deletion mapped, ruled out any involvement of coding sequences in the rearrangement. PCR amplification of the as yet unsequenced murine c-fes IVS3 and IVS4 showed they are about 50% longer than their human and feline homologs. Sequencing of IVS4 showed no difference between tumor and control DNA. Sequencing of part of the approximately 2600-bp IVS3 was guided by the restriction analysis of PCR products from control and hybrid DNAs. This showed that differences from the control appeared to be mainly located in the 900-bp HindIII-EcoRI fragment, localized in the middle of IVS3. As all three hybrids had the same restriction map, this fragment was sequenced in one of them (ESb). A run of >200 CA repeats was found in control DNA, and a reduction in the CAn microsatellite accounted for most of the c-fes deletion in the ESb hybrid. Interestingly, the 50% reduction in the size of human and feline c-fes IVS3 as compared with the murine homolog is mostly due to contraction of the same microsatellite.
Animals; Sequence Homology, Nucleic Acid; Exons; Humans; Protein-Tyrosine Kinases; Mice; Proto-Oncogenes; Polymerase Chain Reaction; Microsatellite Repeats; Base Sequence; Proto-Oncogene Proteins c-fes; Proto-Oncogene Proteins; Tumor Cells, Cultured; Blotting, Southern; Lymphoma, T-Cell; Restriction Mapping; Cats; DNA Primers; DNA, Satellite; Molecular Sequence Data; Species Specificity; Cell Line; Sequence Deletion
Settore MED/03 - Genetica Medica
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/188680
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