Document Type : Research Paper

Authors

• Eskandar Hosseinnezhad Lazarjani ¹
• Abbas Doosti ²
• Ali Sharifzadeh ³

¹ Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran

² Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

³ Department of Microbiology, Faculty of Veterinary Medicine, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

Abstract

In this experimental study, the gene encoding the protein Chapron biogenesis fimbriae Csu was amplified by carbapenem-resistant Acinetobacter baumannii by PCR. the gene was cloned and subcloned into T and pcDNA3.1 (+) vectors, respectively. Confirmation of gene cloning was evaluated by three methods: PCR, cleavage enzymes, and sequencing. Then, 100 μl of 100 ng/ml concentration of the final recombinant vector pcDNA3.1 (+) - csuC was injected into the quadriceps muscle of BALB / c mice and the expression of the target gene in the quadriceps muscle of mice was examined by RT-PCR and the results of expression differences at significant levels P <0.05, P <0.01, and P <0.001 reported. Observation of 831 bp band after RT-PCR in quadriceps muscle of mice in pcDNA3.1 (+) - csuC group compared to control group (PBS) confirms the expression of csuC gene in quadriceps muscle of mice. The results of the study of changes in gene expression in the target group compared to the control group showed that in the target group the expression of the target gene after 2 days showed a significant increase in expression. This increase in expression reached its highest level by day 30 after injection and the difference in expression was significant.

The recombinant pcDNA3.1 (+) - csuC construct made in this study can express the csuC gene of Acinetobacter baumannii in the quadriceps muscle of mice. Also, the pcDNA3.1 (+) - csuC construct has the potential to be considered as a gene vaccine in future research.

Keywords

• Acinetobacter baumannii
• csuC
• recombinant pcDNA3.1 (+) - csuC
• RT-PCR

Main Subjects

• Biotechnology