Jawad Kadhim Sallal Al-Jorani; Mohammadreza Nassiry; Ali Javadmanesh
Abstract
Today, ostrich breeding has been widely developed in Iran and other countries due to the ability of this animal to produce quality meat, leather, and oil. However, one of the main problems in breeding them is sex determination using aggressive techniques with low accuracy. This study aimed to determine ...
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Today, ostrich breeding has been widely developed in Iran and other countries due to the ability of this animal to produce quality meat, leather, and oil. However, one of the main problems in breeding them is sex determination using aggressive techniques with low accuracy. This study aimed to determine the sex of immature ostriches using specific primers in a multiplex PCR reaction. This study considered 20 specimens of unspecified immature and six specimens (three adult males and females) of known-sex African ostriches as controls. SS and OSFES primers were used to amplify part of the female-specific sequence and 18S primer was used as a control in a PCR reaction. The presence of SS and OSFES bands in gel electrophoresis indicated the amplification of the desired parts related to the female sex and the absence of these bands indicates the male sex of the species. In total, out of 20 African ostriches studied, 50% of them belonged to females and 50% of them belonged to males. Later, with the growth of immature individuals, the results of this experiment were confirmed. In this study, it was found that the use of feather samples for DNA extraction and multiplex PCR is a suitable, accurate, and cost-effective method in identifying and determining the sex of young ostrich and leads to more real and reliable results, avoiding stress in birds.
Mohammadreza Nassiri; Azadeh Safarchi; Masoume Vakili-Azghandi; Vinod Gopalan; Mohammad Doosti; Shahrokh Ghovvati; Ahmad Reza Movassaghi
Abstract
p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic ...
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p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic and normal mammary gland of canine as an animal model. Elleven benign and malignant specimens and 5 normal specimens were collected. After RNA extraction and cDNA synthesis, relative quantification of p53 and VEGF-C genes were accomplished by Real-time quantitative PCR (RT-qPCR) based on use of β-actin as a reference gene. The relative mRNA expression of the p53 and VEGF-C genes were analyzed by GLM procedure of SAS software v9.2. The results indicated that the VEGF-C and p53 mRNA expression in neoplastic specimens was over-and down-expressed respectively as compared with normal specimens and p53 mRNA expression was significantly negatively associated with VEGF-C (~4 fold) in neoplastic specimens (P <0.01). The findings emphasized that simultaneous evaluation of p53 and VEGF-C expression can be used as tumor biomarker for early diagnosis of malignancy in canine. Furthermore, RT-qPCR is a rapid and sensitive method to for monitoring and investigating of suspicious canine at the beginning stage of malignancy and may provide an alternative explanation for deregulated p53 signalling in breast cancer.
Fereshteh Ashrafi; Mohammadreza Nassiri; Seyed Abdolrahim Rezaee; Ali Javadmanesh
Abstract
Bovine leukemia virus (BLV) is the etiologic agent of enzootic bovine leucosis (EBL) for the bovine host. In this study to examine gene expression changes in the manifestation of the EBL malignancy, four pooled RNA samples (three RNAs in each sample) were applied for transcriptome sequencing using RNA-seq ...
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Bovine leukemia virus (BLV) is the etiologic agent of enzootic bovine leucosis (EBL) for the bovine host. In this study to examine gene expression changes in the manifestation of the EBL malignancy, four pooled RNA samples (three RNAs in each sample) were applied for transcriptome sequencing using RNA-seq technique. Differential expression analysis was done to compare the infected bovine group with the healthy bovine group using DESeq2 package in R software. Furthermore, functional gene ontology (GO) term and KEGG pathway enrichment analysis were stablished using the DAVID online database to identify involved GO terms and pathways in the host response to BLV infection. Our results suggested that 371 up- and 72 downregulated genes were involved in EBL with statistically significant threshold log2foldchange (LFC) = 1 and false discovery rate (FDR) <0.05 that were enriched in 74 biological processes and 20 KEGG pathways. Most of identified genes were associated with cancer, especially B-cell malignancies. The glycolysis/glycogenesis metabolic process is activated in B cells that confers growth and survival advantages in tumor and dysregulated CXCL10, IL17R, BTK, CDK4 and SYK genes known as valid biomarkers to increase the proliferation of malignant cell. The outcomes can provide a list of involved genes in the malignancy and help to screen candidate genes for cancer therapy in the future.
Ayeh Sadat Sadr; Mohammadreza Nassiri; Seyed Alireza Salami; Mohammad Reza Bakhtiarizadeh; Mojtaba Tahmoorespur; Alireza Shafeinia
Abstract
The poultry industry occupies an important position in the provision of animal protein. Recently, next generation sequencing technology (RNA-Seq) has become available as a powerful tool to investigate transcriptional profiles for gene expression analysis of many organisms. The main use of RNA-Seq in ...
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The poultry industry occupies an important position in the provision of animal protein. Recently, next generation sequencing technology (RNA-Seq) has become available as a powerful tool to investigate transcriptional profiles for gene expression analysis of many organisms. The main use of RNA-Seq in agriculture species are focusing on finding the immune related genes or pathways by comparison of the whole transcriptome following pathogen challenge. Alternative splicing (AS) is the major fundamental mechanism generating the protein diversity and regulating the gene expression in eukaryotic organism. Identifying genes that are differentially spliced between two groups of RNA-sequencing samples is interesting subject in transcriptome with next-generation sequencing technology in this study used RNA sequencing to comparison isoforms of two breeds. A total of 64,819 transcripts were identified by aligning sequence reads to genome among the evaluated isoforms for expression analysis, 310 were significantly differentially expressed between two breeds, including 251 up-regulated and 59 down-regulated. The KEGG results of up regulated isoforms showed that that no pathway was found significantly different (FDR ≤ 0.05). However, enrichment analysis suggested that seven were over-represented (P-value ≤ 0.05) within the up regulated isoforms. Only one of them functionally related to immune system, natural killer cell mediated cytotoxicity. The results showed genes which are breed-specific expression and the comparative transcriptome analysis help to understand the difference of genetic mechanism.
Zahra Roudbari; Mohammadreza Nassiri; Mojtaba Tahmoorespur; Aliakbar Haddad-Mashadrizeh; Ali Javadmanesh
Abstract
Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant ...
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Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant pharmaceutical proteins. In this study, we constructed a lentiviral vector carrying coding region of human GH1 (hGH) gene in order to production of recombinant hGH in mammalian cell line. hGH gene was amplified from a plasmid containing full-length hGH coding sequence and then cloned into the lentiviral vector pCDH-GFP. The HEK293T cells were transduced by the lentivirus particles as a targeted cell. hGH expression status in the recombinant cells were confirmed by RT-PCR. Additionally, western blotting analysis results showed that the recombinant cells maintained a stable hGH expression during five weeks of continuous culture. In conclusion, results of current study suggested that constructed lentiviral vector can potentially be used for a stable production of recombinant hGH protein in HEK293T cells. This methodology could be served as a foundation for further research and may open new insights toward therapeutic protein manufacturing.
