Abnormal splice in a mutant human beta-globin gene not at the site of a mutation.
- 1 March 1983
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 80 (5) , 1184-1188
- https://doi.org/10.1073/pnas.80.5.1184
Abstract
The expression of a cloned mutant human .beta.-globin gene was studied in tissue culture cells. The gene, which was previously isolated from the chromosomal DNA of an individual with a low level of normal .beta.-globin expression (.beta.+-thalassemia), contains 5 mutations inside the large intervening sequence (IVS2), as well as a silent change in codon 2. This .beta.-thalassemia gene (thal) was inserted into a plasmid that is replicated and transcribed in a line of monkey kidney cells in culture. S1 nuclease mapping of the .beta.-globin RNA transcribed from this gene indicates that some of the .beta.-globin RNA is spliced abnormally by using a cryptic 3'' splice sequence normally present in IVS2 but not used in processing the normal .beta.-globin transcript. The cryptic 3'' splice site is not the site of a mutation in the thal gene. Because neither the 5'' or 3'' splice junction nor the cryptic site is mutated in this gene, it is most likely that the mutation at position 705 of IVS2, the only nonpolymorphic change in the gene. Because neither the 5'' or 3'' splice junction nor the cryptic site is mutated in this gene, it is most likely that the mutation at position 705 of IVS2, the only nonpolymorphic change in the gene, interferes indirectly with normal processing. Certain sequences within IVS must be conserved to prevent abnormal splicing and loss of gene function.This publication has 28 references indexed in Scilit:
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