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C.M. Thaung, L. McKie, K. West, J. Morgan, J. Hunter, S.D. Brown, I.J. Jackson, S.H. Cross; Mouse Retinal Degeneration: Seven New Mutations in Pde6b . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4530.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose The Harwell ENU Mouse Mutagenesis Project has generated seven new recessive alleles of the Pde6b gene which cause retinal degeneration. Three of these (called Pde6batrd1-3) show slowed onset degeneration when compared to the classic Pde6brd1 mutant behaviourally, clinically and histologically; while the remaining four (called Pde6brd1-1H - rd1-4H) show a rate of degeneration indistinguishable from that of Pde6brd1. The aim of this work is to characterise the genetic mutations in each of these new alleles. Methods For each mutation, DNA from heterozygous animals (new mutation/ Pde6brd1) was used for capillary-based conformation-sensitive gel electrophoresis. Test regions were amplified by PCR, denatured and renatured and run on an ABI 310 under semi-denaturing conditions. This revealed whether there were any sequence differences between the new mutant and Pde6brd1. If a difference was detected, PCR products from that region were directly sequenced. Results For all seven mutants, the mutation was successfully detected. Missense mutations were found in two of the slow-onset retinal degenerations, and the third was a splice site mutation. Nonsense mutations were found in three of the rapid-onset retinal degenerations and the fourth was a splice site mutation. Conclusions We have successfully detected the genetic mutation for seven new phenotypically abnormal alleles of Pde6b. Two out of three slowed-onset phenotypes have missense mutations, implying that there is residual functional protein. The splice site mutation in the third results in some, but not all, of the transcript being misspliced. Three out of four rapid-onset phenotypes have nonsense (i.e. truncation) mutations, probably resulting in complete loss of function. The splice site mutation in the fourth is in the invariant GT and is predicted to cause a splicing defect leading to the production of an aberrant transcript. The missense mutations in particular will give further information into the structure-function relationship of the Pde6b gene.
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