Molecular Genetics and Genetic Engineering
Samaneh Bagheri; Barat-Ali Fakheri; Ali Mohammad Ahadi; Abbasali Emamjomeh
Abstract
Wheat streak mosaic virus (WSMV) is one of the most important wheat infectious which its prevalence is increasing in Iran. NIa protein as a key protein in WSMV, plays important roles in viral replication and proteolytic digestion of viral polyprotein. Considering the critical role of NIa protein in viral ...
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Wheat streak mosaic virus (WSMV) is one of the most important wheat infectious which its prevalence is increasing in Iran. NIa protein as a key protein in WSMV, plays important roles in viral replication and proteolytic digestion of viral polyprotein. Considering the critical role of NIa protein in viral infection, isolation, sequencing and determination of secondary and tertiary structure of this protein were the objectives of the present study. Also, the amino acids present in the NIa protein active site, which can be used as the targets for design and developement of new antiviral agents, were investigated. In this study, the gene encoding NIa protein was isolated from WSMV and its sequencing was done followed by cloning in pEST prokaryotic vector. The resulted nucleotide sequence was deposited in NCBI database with accession number MK335432. Investigation of physical and chemical properties showed that NIa protein is including 426 amino acids, 48.8 kD molecular weight and 8.81 PI. Prediction of secondary structure of NIa protein revealed that 53.29% of its structure composed of irregular loops, which was justified by the structural dirsorder of this protein. The results of the active site investigation of NIa protein based on homologous sequences alignment and three-dimensional structure of the protein showed a highly conserved site which was included of histidine, aspartic acid and cysteine amino acids.
Molecular Genetics and Genetic Engineering
Hadi Karimbeigi; Farhad Nazarian Firouzabadi; Saber Golkari; Ahmad Ismaili; Masoud Alirezaei
Abstract
Histamine, an important biogenic amine, is produced by many organisms and play a diverse role in living organisms. Many micro-organisms synthesis and stored histamine in food, especially in canned food products, posing health threat to human. Diamine oxidase or histaminase catalysis histamine in healthy ...
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Histamine, an important biogenic amine, is produced by many organisms and play a diverse role in living organisms. Many micro-organisms synthesis and stored histamine in food, especially in canned food products, posing health threat to human. Diamine oxidase or histaminase catalysis histamine in healthy individuals and reduces its harmful side effects. In this study, a full-length cDNA encoding an amine oxidase was isolated from a chickpea (Cicer arietinum) landrace (grit), deposited in Genbank database (KU058599) and cloned in a binary expression vector. The cloned cDNA had an ORF with 2013 bp length encoding a protein with 670 amino acids and a molecular mass of 75.7 KDa. Multiple sequence alignments analysis showed that the active site and important amino acids are highly conserved in E.coli, pea and grit histaminases. Bioinformatics analysis revealed differences in 4 amino acids between grit histaminase and that of GenBank deposited Cicer arietinum amine oxidase (CAA08855). Phylogenetic analysis grouped grit histaminase (KU058599) with plant, especially legume histaminases with 99% bootstrapping.
Molecular Genetics and Genetic Engineering
Masoud Tohidfar; Ebrahimi Ghorishi; Bartali Fakheri; Motahareh Mohsenpour
Volume 4, Issue 6 , October 2014, , Pages 47-59
Abstract
Abiotic stresses such as drought and salinity are the first factor of YIELD decrease in the world. In this study, betaine aldehyde dehydrogenase gene (badh) is used as both an abiotic stress marker gene and one of the abiotic stress tolerance candidate genes along with Flavodoxin (fld) in construction ...
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Abiotic stresses such as drought and salinity are the first factor of YIELD decrease in the world. In this study, betaine aldehyde dehydrogenase gene (badh) is used as both an abiotic stress marker gene and one of the abiotic stress tolerance candidate genes along with Flavodoxin (fld) in construction of chloroplast vector for rice. Thereby, adding the appropriate enzyme role to let gene cassette enter to the center. The first two parts of a specific target areaof Plastom rice (FR) the rice genome using PCR that they can be re-attached in cloning. Then, the gene cassetes were designed for fld and badh genes in regulatory regions of the chloroplast. So that the fld genes and with rbcl 5'UTR and badh genes with T7gene10 5'UTR were cloned the strong chloroplastsic promoter Prrn and the terminator rbcl 3'UTR. Finally, The complete two-gene cassettes fld/badh with the regulatory regions is separated from targeting chloroplast metabolism of rice in two sides cloned. The two specific chloroplast vector called pFrFB(-) and pFrFB(+) are potential to be attached to gene that are resistant to drought and salinity targeted with two different orientations relative to the inner regions of the chloroplast genome of rice plants and are able to be the goal of creating high resistance to salinity, drought and chill in transferring genes to use the chloroplast of rice plant via gene gun.
