Study of the RNA splicing defect in the common Chinese beta-thalassemia gene, IVS-II nt. 654 C-->T by using mRNA/PCR.
With direct sequencing of the amplified cDNA, we analysed the transcript and mRNA splicing defect in a common Chinese beta-thalassemia mutant (IVS-II nt. 654 C-->T). The result shows that this mutant gene would not only produce abnormally processed beta-globin mRNA, but also transcribes a small amount of normally spliced mRNA, hence leading to beta+ thalassemia. The method described herein provides a simple and sensitive approach to the studies of gene expression and molecular defects in genetic diseases at transcriptional level.
Huang
S Z
SZ
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, PRC.
Ren
Z R
ZR
Zeng
Y T
YT
Zeng
F Y
FY
Rodgers
G P
GP
Schecter
A N
AN
D000595:Amino Acid Sequence; D044466:Asian People; D002675:Child, Preschool; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D015870:Gene Expression; D005796:Genes; D005914:Globins / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D017086:beta-Thalassemia / Q000235:genetics*
D012333:RNA, Messenger; D005914:Globins; D004247:DNA