[Analysis of USH2A gene mutation in a Chinese family affected with Usher syndrome].

Abstract

OBJECTIVE:

To investigate the disease-causing mutation in a Chinese family affected with Usher syndrome type II.

METHODS:

All of the 11 members from the family underwent comprehensive ophthalmologic examination and hearing test, and their genomic DNA were isolated from venous leukocytes. PCR and direct sequencing of USH2A gene were performed for the proband. Wild type and mutant type minigene vectors containing exon 42, intron 42 and exon 43 of the USH2A gene were constructed and transfected into Hela cells by lipofectamine reagent. Reverse transcription (RT)-PCR was carried out to verify the splicing of the minigenes.

RESULTS:

Pedigree analysis and clinical diagnosis indicated that the patients have suffered from autosomal recessive Usher syndrome type II. DNA sequencing has detected a homozygous c.8559-2A>G mutation of the USH2A gene in the proband, which has co-segregated with the disease in the family. The mutation has affected a conserved splice site in intron 42, which has led to inactivation of the splice site. Minigene experiment has confirmed the retaining of intron 42 in mature mRNA.

CONCLUSION:

The c.8559-2A>G mutation in the USH2A gene probably underlies the Usher syndrome type II in this family. The splice site mutation has resulted in abnormal splicing of USH2A pre-mRNA.

• Authors: Pengcheng Li, Fei Liu, Mingchang Zhang, Qiufen Wang, Mugen Liu
• Journal: Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics (Zhonghua Yi Xue Yi Chuan Xue Za Zhi) Vol. 32 Issue 4 Pg. 468-71 (Aug 2015) ISSN: 1003-9406 [Print] China
• PMID: 26252086 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Extracellular Matrix Proteins
• USH2A protein, human

Topics

• Adolescent
• Adult
• Aged
• Asian People (genetics)
• Base Sequence
• Child
• China
• Extracellular Matrix Proteins (genetics, metabolism)
• Female
• Humans
• Male
• Middle Aged
• Molecular Sequence Data
• Pedigree
• Usher Syndromes (genetics, metabolism)
• Young Adult