Sheffield V C, Fishman G A, Beck J S, Kimura A E, Stone E M
Department of Pediatrics, University of Iowa, Iowa City.
Am J Hum Genet. 1991 Oct;49(4):699-706.
Retinitis pigmentosa (RP) is a group of disorders characterized by progressive degeneration of the outer retina, resulting in night blindness, visual field loss, an abnormal electroretinogram, and characteristic retinal pigmentary changes. An important step in the understanding of RP has been the recognition that some cases of autosomal dominant RP (ADRP) are caused by mutations in the rhodopsin gene. Multiple different point mutations within the coding sequence of the rhodopsin gene have been associated with ADRP. We have developed a GC-clamped denaturing-gradient-gel electrophoresis (DGGE) assay for the coding region of the rhodopsin gene and have used this assay to screen ADRP patients for mutations. The assay consists of amplifying with PCR the five exons of the rhodopsin gene and then analyzing each PCR product by DGGE. We have used this assay to detect three previously unreported rhodopsin base substitutions associated with ADRP. The use of this assay to identify ADRP patients who have various rhodopsin mutations has allowed us to begin studies seeking to correlate molecular genotype with clinical phenotype. Furthermore, GC-clamped DGGE has allowed us to identify families with ADRP not caused by a rhodopsin mutation. Such families will be important in the search for other genes involved in ADRP.
视网膜色素变性（RP）是一组以外侧视网膜进行性退化为特征的疾病，可导致夜盲、视野缺损、视网膜电图异常以及特征性的视网膜色素改变。理解RP的一个重要进展是认识到某些常染色体显性视网膜色素变性（ADRP）病例是由视紫红质基因突变引起的。视紫红质基因编码序列内的多个不同点突变已与ADRP相关联。我们针对视紫红质基因的编码区开发了一种GC夹变性梯度凝胶电泳（DGGE）检测方法，并使用该方法对ADRP患者进行突变筛查。该检测方法包括用PCR扩增视紫红质基因的五个外显子，然后通过DGGE分析每个PCR产物。我们已使用该检测方法检测到三个先前未报道的与ADRP相关的视紫红质碱基替换。使用该检测方法来识别具有各种视紫红质突变的ADRP患者，使我们能够开始进行旨在将分子基因型与临床表型相关联的研究。此外，GC夹DGGE使我们能够识别并非由视紫红质突变引起的ADRP家族。这类家族对于寻找参与ADRP的其他基因将具有重要意义。
