关键词： B3GALTL B3GALTL gene B3GTL CD CV DNA DNA complementary to RNA DNase ESE Exon skipping F Functional analysis G418 Geneticin HSF Ins MMLV Moloney Murine Leukemia Virus NMD PCR PPS PTC Peters plus syndrome PolyA R RPMI RT-PCR Roswell Park Memorial Institute SR TMR WT aa amino acid(s) base pair(s) beta-1,3-glucosyltransferase beta3-glycosyltransferase-like bp cDNA catalytic domain consensus value dNTP deoxyribonuclease deoxyribonucleic acid deoxyribonucleoside triphosphate exonic splicing enhancer forward human splicing finder insertion mRNA messenger ribonucleic acid nonsense-mediated mRNA decay polyadenylation polymerase chain reaction premature translation stop codon reverse reverse transcription polymerase chain reaction splice site ss stem region transmembrane region wild type β-galactosidase βGal

Mesh ： Cleft Lip / genetics Computer Simulation Cornea / abnormalities Exons Galactosyltransferases / chemistry genetics metabolism Glucosyltransferases / chemistry genetics metabolism Growth Disorders / genetics Humans Limb Deformities, Congenital / genetics Mutation Nonsense Mediated mRNA Decay RNA Splice Sites RNA Splicing Tunisia

来  源：   DOI：10.1016/j.gene.2013.07.058   PDF(Sci-hub)

Abstract：
Peters plus syndrome is a rare recessive autosomal disorder comprising ocular anterior segment dysgenesis, short stature, hand abnormalities and distinctive facial features. It was related only to mutations in the B3GALTL gene in the 13q12.3 region. In this study, we undertook the first functional analysis of a novel c.597-2 A>G splicing mutation within the B3GALTL gene using an ex-vivo approach. The results showed a complete skipping of exon 8 in the B3GALTL cDNA, which altered the open reading frame of the mutant transcript and generated a PTC within exon 9. This finding potentially elicits the nonsense mRNA to degradation by NMD (nonsense-mediated mRNA decay). The theoretical consequences of splice site mutations, predicted with the bioinformatics tool Human Splice Finder, were investigated and evaluated in relation to ex-vivo results. The findings confirmed the key role played by the B3GALTL gene in typical Peters-plus syndromes and the utility of mRNA analysis to understand the primary impacts of this mutation and the phenotype of the disease.
摘要：
Petersplus综合征是一种罕见的隐性常染色体疾病，包括眼前节发育不全，身材矮小,手部异常和独特的面部特征。它仅与13q12.3区域中B3GALTL基因的突变有关。在这项研究中,我们使用离体方法对B3GALTL基因内的新型c.597-2A>G剪接突变进行了首次功能分析。结果表明，B3GALTLcDNA中外显子8完全跳跃，它改变了突变体转录本的开放阅读框，并在外显子9内产生了PTC。该发现潜在地引起无义mRNA被NMD降解(无义介导的mRNA衰变)。剪接位点突变的理论后果，用生物信息学工具HumanSpliceFinder预测，进行了与离体结果相关的调查和评估。研究结果证实了B3GALTL基因在典型的Peters-plus综合征中的关键作用，以及mRNA分析的实用性，以了解这种突变的主要影响和疾病的表型。
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