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.
Mahdi Soltani; Mohammadreza Nassiri; Alireza Sadrebazzaz; Mojtaba Tahmoorespoor
Abstract
Neospora caninum is an obligate intracellular parasitic protozoa and considered as causal agent of Neosporosis which infect wide variety of hosts. NcGRA7 is an immunodominant antigen recognized by sera from bovines, naturally infected by N. caninum, which is used as a powerful target for recombinant ...
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Neospora caninum is an obligate intracellular parasitic protozoa and considered as causal agent of Neosporosis which infect wide variety of hosts. NcGRA7 is an immunodominant antigen recognized by sera from bovines, naturally infected by N. caninum, which is used as a powerful target for recombinant or DNA vaccine preparation against neosporosis. There is no study about identifying the molecular structure of Neospora caninum in Iran, so as first step, current study tried to identify NcGRA7 gene in this parasite in Iran. After extraction of total RNA from N. caninum tachyzoites, cDNA was synthesized and NcGRA7 gene was amplified using cDNA as template. Then the PCR product was cloned into pTZ57R/T vector and transformed into Escherichia coli (DH5α strain), and the resulted recombinant plasmid was submitted for sequencing, followed by bioinformatics analysis. The data obtained from sequencing of native NcGRA7 was recorded in GenBank. The deduced amino acid sequence of NcGRA7 in current study was compared with other N. caninum NcGRA7 sequences and showed some identities and differences. NcGRA7 gene of N. caninum was successfully cloned into the pTZ57R/T vector and recombination was confirmed by sequencing, colony PCR, and enzymatic digestion, making it ready expression of recombinant protein for further studies.
Abdul-raoof Al-shawkany; Mohammadreza Nassiri; Saied Zibaei; Mojtaba Tahmoorespour; Sayed Mahdi Ziaratnia; Mohsen Fathi Najafi; Alireza Haghparast; Shahrokh Ghovvati; Seyed Hassan Pourseyed; Mohammad Rashtibaf
Abstract
Foot-and-mouth disease (FMD) is a major cause of endemic outbreaks in livestock in Iran. In this study, clinical field samples of foot-and-mouth disease virus were collected from an outbreak in Khorasan Razavi Province during April and August of 2010, and subjected to indirect sandwich ELISA and RT-PCR. ...
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Foot-and-mouth disease (FMD) is a major cause of endemic outbreaks in livestock in Iran. In this study, clinical field samples of foot-and-mouth disease virus were collected from an outbreak in Khorasan Razavi Province during April and August of 2010, and subjected to indirect sandwich ELISA and RT-PCR. The viral serotype circulating during the period was confirmed to be type O. The virus was then genetically characterized for its complete P1 genomic sequences to compare with nine corresponding nucleotide sequences of representative foot-and-mouth disease viruses (FMDVs) registered in the GenBank. The P1-coding region was 2208 nucleotides in length with 736 encoded amino acid residues. Phylogenetic analysis revealed two major lineages of A (with three additional clusters) and B. Iranian field isolate was grouped within cluster I, most closely related to Pakistani strains PAK/39/2008 and PAK/29/2008 sharing 98.37 and 98.1% amino acid identity, respectively, demonstrating the close epidemiological links between countries in the region. In contrast, our isolate showed low amino acid identity with Italian isolate O-2-Brescia (93.48%) and Argentinean isolate O1 Caseros (93.75%). Based on multiple sequence alignments, comparison of sequences showed that the characteristic amino acid mutations were found in the VP1, VP2 and VP3 proteins of isolated virus. This article is the first to report on the complete P1 genomic characterization of type O FMDV circulating in Iran
