Identification of a MIP mutation that activates a cryptic acceptor splice site in the 3' untranslated region.

ABSTRACT:

Purpose

To investigate the consequence of a major intrinsic protein MIP splice-site mutation (c.607-1G>A) in a four-generation Chinese pedigree afflicted with autosomal dominant congenital cataracts (ADCC).

Methods

Both a mutated minigene with c.607-1G>A, and a wild-type minigene were constructed using the pTARGET mammalian expression vector. They were transiently transfected into HeLa cells and human lens epithelial cells, respectively. After 48 h incubation, RNA extraction and RT-PCR analysis were performed and PCR products were separated and confirmed by sequencing. Structural models of the wild-type and the mutant aquaporin 0 (AQP0) were generated and analyzed using SWISS-MODEL.

Results

The G>A transition activated a cryptic acceptor splice site (c.965-966) in the

SUBMITTER: Jin C 

PROVIDER: S-EPMC2994330 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature