Gaucher disease is the most common lysosomal disease and is highly prevalent in the Ashkenazi Jewish (AJ) population. It is an autosomal recessive disorder and it caused by a deficiency in glucocerebrosidase enzymatic activity due to various mutations in the gene encoding lysosomal glucocerebrosidase (GBA). Glucocerebrosidase is found in the lysosomes of nearly all cell types, and is responsible for the degradation by hydrolysis of metabolic intermediates derived from the cellular turnover of cell membrane (Zuckerman et al, 2007).
Figure 1-Gaucher cells.
Figure 2-The normal sequences and the mutated sequences that used as primers for generating the positive controls.
Figure 3-A. PCR products using six different primers in the absence of and in the presence of a positive control (the plasmid). B. Six mutated primers to amplify IVS2+1 mutation.
Figure 4-A. PCR products using five different primers in the absence of and in the presence of a positive control (the plasmid). B. Five mutated primers to amplify 1226G mutation.
Gaucher disease is an autosomal, recessive, lysosomal storage disease most prevalent in the Ashkenazi Jewish population. Gaucher disease is caused by a deficiency in glucocerebrosidase enzymatic activity, with subsequent accumulation of glucosylceramide in various organs. In the Jewish Ashkenazi population there are four known mutations that cause the disease:1226G (N370S),84GG,IVS2+1 and 1448C (L444P). By using PCR mutagenesis, here we developed an amplification specific PCR protocol for the detection of the presence of two Gaucher mutations in the Ashkenazi Jewish population. Developing an amplification specific PCR protocol for the mutations of Gaucher disease is very important both for detecting carriers and homozygotes for the disease and for determine mutations frequencies.
I would like to thank Alex Bulanov and Xie Xie for their help, infinite patience and support, without which this project would not have been possible. In addition I would like to thank Dr. Berish Rubin for his guidance and providing the opportunity to take a part in this project and to achieve this amazing experience. I would like to thank also Dr. Sylvia Anderson for helping and answering any question I had.
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