Mutational analysis of the glucose-6-phosphate translocase gene involved in the glycogen storage disease type 1b

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Mutational analysis of the glucose-6-phosphate translocase gene involved in the glycogen storage disease type 1b

 

Author: Chan, Bik-yan Selena
Title: Mutational analysis of the glucose-6-phosphate translocase gene involved in the glycogen storage disease type 1b
Degree: M.Sc.
Year: 2000
Subject: Glycogenosis
Glucose-6-phosphatase
Mutation (Biology)
Hong Kong Polytechnic University -- Dissertations
Department: Multi-disciplinary Studies
Dept. of Nursing and Health Sciences
Pages: viii, 51 leaves : ill. (some col.) ; 30 cm
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b1541798
URI: http://theses.lib.polyu.edu.hk/handle/200/84
Abstract: Glycogen storage disease type 1 is caused by the deficiency of microsomal glucose-6-phosphatase system in the liver. More than 80% of patients with glycogen storage disease type I are caused by a deficiency of glucose-6-phosphatase and classified as glycogen storage disease type 1a. The gene for glucose-6-phosphatase was identified and mapped on chromosome 17, and more than 20 mutations have been reported to date. The second common subtype of glycogen storage disease type 1 is type 1b. It is an autosomal recessive disorder caused by a deficiency of glucose- 6-phosphate translocase. In addition to the clinical symptoms of glycogen storage disease type 1a, patients with glycogen storage disease type 1b commonly have infectious complications which are caused by neutropenia and dysfunction of neutrophils. In this project, the glucose-6-phosphate translocase gene of a Chinese patient diagnosed as glycogen storage disease lb was analysed. All the nine exons were directly sequenced to find out the mutation. The allele frequency of the disease mutation was found by screening 351 cord blood samples of normal Chinese individuals by amplification refractory mutation system (ARMS). In addition, the non-coding region of the glucose-6-phospbate translocase gene were amplified using polymerase chain reaction (PCR) and screened for single nucleotide polymorphisms (SNPs) using normal DNA samples by single stranded conformation polymorphism (SSCP) anaylsis. DNA sequencing of all nine exons revealed a homozygous G->A transition at nucleotide position 5131 of the genomic DNA sequence. The missense mutation is loacted in exon 3 at codon 149, changing the non-polar glycine to the charged glutamic acid, i.e. G149E. The missense mutation was confirmed by Bpml restriction analysis. The allele frequency of the G149E mutation was found to be 0.0028. In addition, 3 common polymorphisms were found in the non-coding region of the glucose-6-phosphate translocase gene. One polymorphism was located in the 5' untranslated region and 2 polymorphisms were located in the 3' untranslated region. For the polymorphism located in the 5' untranslated region, nine different alleles were found. The allele frequency of the most common allele was 0.7824 and the rest ranging from 0.0118 to 0.0647. For the polymorphism located in fragment 1 of the 3' untranslated region, two different alleles were found with the allele frequencies of 0.7952 and 0.2048. Direct sequencing result on these alleles revealed a C->G homozygous base substitution at nucleotide 8153. For the polymorphism located in fragment 4 of the 3' untranslated region, two different alleles were found with allele frequencies of 0.8967 and 0.1033. The direct sequencing result on these alleles revealed a homozygous TT deletion at nucleotide 8907-8908. In conclusion, the G149E mutation may be a common cause of glycogen storage disease 1b in the Chinese population.

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