小基因
外显子
囊性纤维化跨膜传导调节器
RNA剪接
桑格测序
分子生物学
生物
内含子
突变
遗传学
基因
编码区
信使核糖核酸
囊性纤维化
选择性拼接
核糖核酸
作者
Xinyue Zhao,Keqiang Liu,Wenshuai Xu,Meng Xiao,Qianli Zhang,Jiaxing Song,Keqi Chen,Yaping Liu,Xinlun Tian,Kai–Feng Xu,Xue Zhang
标识
DOI:10.1007/s11684-021-0846-5
摘要
Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
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