Showing 5 results for Sscp
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Volume 8, Issue 2 (10-2017)
Abstract
Introduction: Hypertrophic cardiomyopathy (HCM) is a rare cardiac disorder which can result in sudden death in young people. Molecular studies have been showed that mutations in the beta-myosin heavy chain (MYH7) gene are one of the most common genetic causes of HCM. The aim of this research was to study exons 8, 9, and 30 of MYH7 gene for possible mutation in HCM patients from Chahar Mahal va Bakhtyari Province.
Material and Methods: In this descriptive-laboratory study, DNA was extracted from 27 blood samples by phenol-chloroform method. DNA samples were then used for PCR-SSCP analysis for amplification and identification of mutation. The Suspected cases with possibility of having mutation were sequenced and the results were observed by Chromas software.
Results: 7 suspected cases recognized by PCR_SSCP were sequenced with forward and reverse primers to verify the presence of mutation.
Conclusion: Mutations in these exons do not have a role in establishment of disease in the studied population. However, study greater number of HCM cases and other exons of this gene are recommended to find the relationship between gene and HCM and to gain necessary information for treatment and management of disease.
M.r. Kalbassi , E. Jorfi , M. Sadeghizadeh, S. Amirinia,
Volume 9, Issue 4 (12-2018)
Abstract
Aims: Continuous monitoring of aquatic genetic diversity among different populations in fish hatcheries is an essential requirement to maintain the viability and sustainability of aquaculture industry. The aim of this study was cloning, sequencing, and detection of major histocompatibility complex (MHC) class II β in silver carp.
Materials and Methods: In this experimental study, the polymorphism of MHC class II β in 138 species of silver carp was studied in 4 different hatcheries of Iran (Guilan, Mazandaran, Golestan, and Khuzestan provinces) in addition to an imported group from China. By polymerase chain reaction (PCR), Hymo-DAB gene amplification was performed and the different haplotypes of the samples were analyzed by single-strand conformation polymorphism (SSCP) method and the sequences obtained with ClustalW2 were matched in Geneious 4.8.5 software and the phylogeny tree of the sequences was plotted.
Findings: The PCR reaction of the MHC-DAB II genome of the silver carp with a weight of about 350bp without side band was obtained in the samples, indicating the amplification of t Hymo-DAB1*01/DAB2*1 gene in silver carp. The highest and lowest diversity of haplotypes was observed in populations of Khuzestan and Mazandaran. The mean difference between synonymous site (dS) and nonsynonymous site (dN) of alleles was 0.25 and 0.30, respectively, with the ratio of 1.2. The highest allelic richness was observed in samples imported from China (5) and the lowest allelic richness was among Mazandaran species (3.8).
Conclusion: Haplotype diversity in silver carp belongs to Hymo-DAB1*01/DAB2*1 gene and among different groups of this species, the highest haplotype diversity is in the Khuzestan population and the highest allelic richness is related to samples imported from China.
Volume 14, Issue 3 (5-2012)
Abstract
Calpastatin (CAST) is a specific inhibiter of calpains, playing a role in meat tenderization and myogenesis. In the present study, the polymorphism of the CAST gene of Makoei sheep was investigated by polymerase chain reaction and single strand conformation polymorphism technique (PCR–SSCP). Genomic DNA was extracted from whole blood samples collected from 100 sheep. A 622 bp CAST exon 1 segment was amplified by standard PCR, using the locus specific primers. PCR products were subjected to a non-denaturing gel electrophoresis. Four SSCP patterns, representing four different genotypes, were identified. The frequencies of the observed genotypes were 0.31, 0.04, 0.63 and 0.02 for AA, BB AB and AC, respectively. Allele frequencies were 0.6313, 0.3586 and 0.01 for A, B and C, respectively. The Observed heterozygosity (Hobs) value for CAST gene was 0.4728. The chi-square test showed significant (P< 0.01) deviation from Hardy-Weinberg equilibrium for this locus in Makoei sheep population.
Volume 15, Issue 6 (11-2013)
Abstract
Baluchi sheep is considered as the most common breed in Iran, constituting about 30% of total sheep population, approximately 15 million heads. This research was designed to study the incidence of mutation in two loci of IGF-I (Exon 3) and ADRB3 (Intron) along with their association with body weight traits in Baluchi sheep population. Following DNA extraction from 190 Baluchi sheep, two pairs of primers were designed to amplify each gene. PCR-SSCP (Single-Strand Conformation Polymorphism) and DNA sequencing were employed to detect polymorphism of the mentioned genes. Two banding patterns were detected for IGF-I locus. The frequencies of AA, AB were recorded as 0.89, 0.11, respectively in IGF-I locus. As for ADRB3, two patterns corresponding with two genotypes (their frequencies mentioned in parentheses) of AA (0.85) and AB (0.15) were identified. One SNP change was observed in ADRB3, and one in IGF-I. A previously reported SNP was detected in exon 3 of IGF-I. The effects of IGF-I and ADRB3 polymorphism on the corrected phenotypes for body weight at birth (BW), weaning (WW), 6 months of age (6MW), 9 months of age (9MW) and 12 months of age or Yearling Weight (YW) were examined using least square methods. No significant association was detected between the polymorphism of IGF-I and body weights. As for ADRB3, the genotype AA was found out to exert a significant positive effect on 6MW (AA, 30.20±1.85 kg day-1; AB, 27.67±1.98 kg day-1; P<0.05).
Volume 20, Issue 1 (6-2017)
Abstract
Objective: Familial hypercholesterolemia (FH) is an autosomal dominant disease mainly attributed to mutations in the low-density lipoprotein receptor gene (LDLR). This study aims to investigate molecular changes in the LDLR gene in patients with high cholesterol in individuals from Ardebil Province, Iran.
Methods: We evaluated 100 patients with suspected FH from Ardebil Province. DNA samples using primers LDLR gene and exon 10 PCR-SSCP method was tested and modified bands on gel electrophoresis detected and subsequently examined by DNA sequencing.
Results: We evaluated 100 patients with suspected FH, 43 males and 66 females. The average age of 50.68 and 281.81 had average cholesterol levels of the subjects. In this study, we identified a polymorphism 1413G(A LDLR gene. Allele G, 70% of the population studied and the A allele is 30% of the subjects to be included.
Conclusion: The findings of this study showed that polymorphism of the LDLR gene in FH 1413GA was not the main role, but could indirectly affect, and possibly other exons of the gene or other genes in the development of FH in the region have.
