1- MSc Student of Cellular and Molecular Biology, Department of Cell Biology, Faculty of Science, Imam Hossein University, Tehran, Iran 2- PhD of Bacteriology, Department of Cell Biology, Faculty of Science, Imam Hossein University, Tehran, Iran , m_saadati@yahoo.com 3- MSc Student of Cellular and Molecular Biology, Islamic Azad University, Central Tehran Branch, Young Reserchers Club, Tehran, Iran 4- PhD of Microbiology, Department of Industrial and Environmental Biotechnology, National Institute of Genetics and Biotechnology, Tehran, Iran 5- PhD of Molecular Genetics, Department of Industrial and Environmental Biotechnology, National Institute of Genetics and Biotechnology, Tehran, Iran 6- Young Researchers Club, Islamic Azad University, Science and Research Campus, Tehran, Iran 7- Researcher, Department of Cell Biology, Faculty of Science, Imam Hossein University, Tehran, Iran 8- PhD of Molecular Genetics, Islamic Azad University, Tehran Medical Branch, Tehran, Iran
Abstract: (16379 Views)
Background: Shigellosis disease is the major causes of morbidity in children with diarrhea in Iran. The virG (icsA) gene plays a key role in pathogenesis and ability of invasion in shigella. The aim of this study was cloning, sequencing virG gene and developing a mutant construct pGEM∆virG in order to induction recombination in a native shigella for generation a live attenuated vaccine candidate strain.
Materials and methods: Initially, by use of biochemical tests, the native shigella strain was detected. The virG gene was cloned in pGEM-7zf vector and the nucleotide sequence was determined. According to the data of sequencing, digestion mapping of pGEMvirG vactor was obtained and a part of virG gene by using enzymatic digestion was removed. Finally, pGEM∆virG construct was transformed to E. coli by utilization of chemical transformation method.
Results: The native shigella strain by using biochemical tests was confirmed. The result of sequencing virG gene (native strain) was submitted in NCBI Genebank database. The pGEM∆virG construct contains a mutant construct of virG gene which 1751 bp was deleted through enzymatic digestion reaction and transformed in E. coli. Conclusion: Using the technique of allelic exchange based on the incident of recombination in bacteria is one of the most effective methods to develop a disruption in the target genes. This mutant construct can be applied in development of a live attenuated Shigella dysenteriae vaccine candidate.
Ahmadi Danesh H, Saadati M, Doroudian M, Yakhchali B, Karkhaneh A A, Hosseini S M, et al . Cloning virG gene and generation mutant construct of pGEM∆virG in order to induction recombination in native Shigella dysenteriae . MEDICAL SCIENCES 2012; 22 (3) :184-190 URL: http://tmuj.iautmu.ac.ir/article-1-590-en.html