HEMOGLOBIN ALPHA 1 (HBA1) GENE SEQUENCE ANALYSIS IN THE THALASSEMIA AFFECTED PATIENTS
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Author:
HEEFA SARWAR
Citable URI :
https://vspace.vu.edu.pk/detail.aspx?id=365
Publisher :
Virtual University
Date Issued:
7/6/2020 12:00:00 AM
Abstract
Alpha-thalassemia is an inherited blood disorder which is an autosomal recessive type
disorder characterized by a microcytic hypochromic anemia and hemolytic anemia. The
a-thalassemias involve the genes HBA1 and HBA2.The aim of this research was to determine
the mutations in hemoglobin alpha 1 (HBA1) in Thalassemia affected patients and in silico
analysis of identified mutations to predict the functional effect. In this study, genomic DNA
was extracted from 40 Patients affected with Thalassemia (n=40) disease. Blood samples
were collected in vacutainers with EDTA as an anticoagulant from the patients and relatives.
Blood samples with anticoagulant were used for leukocytes based DNA extraction. Standard
organic method was used for DNA extraction. DNA samples were quantified using agarose
gel and DNA ladder. Primers were designed using gene sequence from NCBI gene bank.
Primer3 software was used for primer designing. PCR conditions will be optimized for
amplification and PCR was performed to determine the SNPs. A 382 base pair fragment of
DNA of HBA1gene of exon 3 was amplified using polymerize chain reaction (PCR)
technique. Sanger sequencing of the selected samples was done to identify polymorphisms.
A total of 24 samples out of 40 samples of DNA were sequenced and these SNPs were
confirmed by alignment. We were unable to find the mutations in the HBA1 gene but two
heterozygous variations were found in HBA1exon 3.Two
heterozygous
variations were confirmed in exonic area of HBA1 gene of
Patients affected with Thalassemia. The findings of this
research revealed no mutations were found in HBA1 gene. Two
heterozygous variants were confirmed at the position of c.514
on amplified fragment from G> C and second change at the
position of c.470 on amplified fragment G > C in 3’UTR..
Variations were further subjected for splice site analysis.
The splicing site analysis was done by using an online tool
(Human splicing finder).A variation which were present at
c.514 G>C were found in the potential splice site and its
consensus value is 88.39 and the second one is not in the
target region of splicing.
URI :
https://vspace.vu.edu.pk/details.aspx?id=365
Citation:
SARWAR ,H(2019),HEMOGLOBIN ALPHA 1 (HBA1) GENE SEQUENCE ANALYSIS IN THE THALASSEMIA AFFECTED PATIENTS,VIRTUAL UNIVERSITY OF PAKISTAN , (Lahore,Pakistan).
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Final Version
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