Identification and characterisation of human dysferlin transcript variants: implications for dysferlin mutational screening and isoforms.
In conducting dysferlin mutational screening using blood mRNA instead of genomic DNA, we identified the occurrence of alternative splicing involving novel dysferlin exons, i.e. exons 5a and 40a, in addition to previously reported alternative splicing of exon 17. Further study employing long range RT-PCR and subcloning revealed a total of fourteen dysferlin transcripts with maintained dysferlin reading frame. The study also characterised the differences in relative frequencies of the dysferlin transcripts in skeletal muscle and blood. The findings have potential implications for molecular diagnosis of dysferlinopathy and the identification of dysferlin isoforms.
Pramono
Zacharias Aloysius Dwi
ZA
Department of Clinical Research, SingHealth Research Facility, Block C, #03-3, Singapore General Hospital, 9 Hospital Drive, Singapore, 169612, Singapore. dpsarengat@nsc.gov.sg
Tan
Chin Lai
CL
Seah
Irene Ai Lian
IA
See
Joseph Shean Long
JS
Kam
Siok Yuen
SY
Lai
Poh San
PS
Yee
Woon Chee
WC
D017398:Alternative Splicing; D001483:Base Sequence; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D049310:Distal Myopathies / Q000235:genetics; D000073939:Dysferlin; D005091:Exons; D014644:Genetic Variation; D006801:Humans; D007438:Introns; D008565:Membrane Proteins / Q000235:genetics*; D009124:Muscle Proteins / Q000235:genetics*; D018482:Muscle, Skeletal / Q000378:metabolism; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger