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Study Of Genetic Polymorphism In Exon 7 And 9 Of Glucosidase Beta Acid (Gba) Gene In Gaucher Diseased Patients From Punjab, Pakistan

by Ayesha Khalid (2013-VA-07) | Dr. M. Yasir Zahoor | Dr. Sehrish Firyal | Mr. Tariq Mahmood.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Gaucher disease (GD) is an inborn metabolic disease transmitted through recessive pattern of inheritance and it is a pan-ethnic disease. It is the most common lysosomal storage disease caused by the deficiency of glucocerebrosidase (GCase), a lysosomal enzyme use in the degradation of macromolecules into simpler molecules. Glucosidase beta acid (GBA) gene encode glucocerebrosidase enzyme and mutations in this gene is responsible for glucocerebrosidase deficiency which results in an accumulation of unbroken glycolipids in those organs rich in monocyte-phagocyte immune system elements i.e. spleen, liver, bone marrow and leads to histological changes. GBA is located on chromosome 1q21 consisting of 11 exons and 10 introns having 7.8kb length. It is divided into three types (I, II and III) on the basis of neurological involvement. More than 300 mutations have been reported in GBA and cause the GD. The present study was performed in order to characterize GBA gene in GD patients from Punjab. Blood samples of 10 patients,enrolled in Children Hospital, Lahore, were taken from DNA repository of Molecular and Genomic Lab at IBBT, UVAS Lahore. The DNA was extracted using organic method. Next step was the amplification of extracted DNA using PCR. After it, the PCR product is purified and this purified PCR product was sent for sequencing. Sequencing of exon 4, 7 and 9 was done using dideoxy sequencing method. After applying different bioinformatics tool, it was found that there was no muttaion in these exons but a heterozygotic variation G>A was found in intron 8. This finging will help in demonstration of molecular pathogenesis of Gaucher disease. Availability: Items available for loan: UVAS Library [Call number: 2338-T] (1).

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Expression And Purification Of A Potent Surface Antigen (Sag1) Of Toxoplasma Gondii In Prokaryotic Expression System

by Zunaira Zafar (2009-VA-542) | Dr. Wasim Shehzad | Dr. M. Yasir Zahoor | Dr. Imran Rashid.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Toxoplasma gondii, an intracellular obligate parasite infects almost all warm-blooded animals including human. Toxoplasmosis, caused by T. gondii, may show minute to severe clinical results in humans. Currently, there is no vaccine available for human use. SAG1 is a major candidate of interest for vaccine development that elicits humoral as well as cellular immune response against this devastating parasite. rSAG1 that had already been ligated in pET28/His expression vector, was transformed in E. coli (BL21) host and expression was confirmed by means of SDS-PAGE and western blotting. Nickel columns were utilized for affinity based chromatographic purification of rSAG1. This purified protein was then quantified via protein quantification kit. Immunogenic recombinant SAG1 can be used in diagnostic antigen-antibody tests e.g. in ELISA. Moreover, it might be used in vaccination against T. gondii. Vaccine against this parasite may alleviate socio-economic burden on human society that ultimately modulates the health parameters for better living. Availability: Items available for loan: UVAS Library [Call number: 2393-T] (1).

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