Challenging for Expression Bovine Rotavirus (RF Strain) Full-Length VP7 Protein in Prokaryotic System
Journal Title: International Journal of Medical Laboratory - Year 2017, Vol 4, Issue 2
Abstract
Background and Aims: Rotavirus enteritis is an acute viral infectious disease among infants. VP7 protein has a key role in attachment and entry virus into the target cell. The VP7 protein is involved in inducing the production of neutralizing antibodies that protect infants against reinfection of the virus. The aim of this study was to heterologous expression of the VP7 gene of bovine rotavirus as a fusion protein of Trx–VP7 in a genetically engineered bacteria. Materials and Methods: Total RNA was extracted from MA104 cells infected with bovine rotavirus strain RF. BRV VP7 gene was amplified using polymerase chain reaction. Another gene was the VP7 synthetic that made in the pBSK plasmid. The pBSK-VP7 plasmid was digested using BamHI and SacI restriction endonucleases, then recombined into the prokaryotic expression vector pET32a. The pET32a-VP7 amplified and pET32a-VP7 synthetic were transformed into BL21 (DE3) competent cells of Escherichia coli, respectively, and induced by different concentrations of Isopropyl β-D-1-thiogalactopyranoside at 30°C and 37°C in luria bertani and terrific broth media, then analyzed using SDS-PAGE and western blotting. Results: SDS-PAGE analysis showed that the both pET32a-VP7 amplified and pET32a-VP7 synthetic were not expressed in the BL21 (DE3) cells, But the expression of pET32a-VP7 synthetic was weak at 30° C in luria bertani media. Conclusions: This is the first report of the production of bovine rotavirus (RF strain) Full-Length VP7 in prokaryotic system expression. VP7 is a membrane protein and has toxic domains that show high toxicity in this expression system.
Authors and Affiliations
Milad Zandi, Ali Teimouri, Manouchehr Makvandi, Mehdi Parsa, Shahram Jalilian
Keywords
Related Articles
- EP ID EP212462
- DOI -
- Views 148
- Downloads 0
Determination of Helicobacter pylori Antigen in Stool Samples and Comparison with Rapid Urease Test in Patients Suspected of Helicobacter Infection
Background and Aims: Helicobacter pylori (H. Pylori) is the microorganism that infects nearly half of word's population. There are several invasive and non-invasive methods for diagnosis of infection. The main objective...
The Relation of Serum Bilirubin Level with the Severity and Complexity of Coronary Artery Disease and Long-term Outcomes in the Patients Undergoing Primary Percutaneous Coronary Intervention
Background and Aims: Bilirubin has been considered an antioxidant that protects against atherosclerosis. The aim of this study was to evaluate the relationship of serum bilirubin level with the severity and complexity of...
Effects of Thymol on Serum Biochemical and Antioxidant Indices in Kindled Rats
Background and Aims: Kindling is regarded as an experimental model of temporal lobe epilepsy that reflects a process of progressive and persistent intensification of seizure. Experimental and clinical evidence suggest th...
Prevalence of qnr Genes in Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae Isolated from Clinical Urine Specimens in University Teaching Hospitals, Iran
Background and Aims: Extended-spectrum β-lactamase (ESBL) producing gram negative bacteria are resistant to penicillins, narrow and extended-spectrum cephalosporins, and aztreonam; also they are frequently resistant to t...
Evaluation of c-Myc mRNA Expression Level in Benign Prostatic Hyperplasia and Prostatic Adenocarcinoma Tissues and Its Correlation with Clinicopathological Characteristics
Background and Aims: Prostate cancer (PCa) is one of the most common cancers among men in Iran. Since changes in the regulation of proto-oncogenes expression are the main causes of most human cancers, including PCa, eval...