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Paper Information

Journal:   MEDICAL SCIENCES JOURNAL OF ISLAMIC AZAD UNIVERSITY   FALL 2012 , Volume 22 , Number 3 (69); Page(s) 184 To 190.
 
Paper: 

CLONINGVIRG GENE AND GENERATION MUTANT CONSTRUCT OF PGEM VIRG IN ORDER TO INDUCTION RECOMBINATION IN NATIVE SHIGELLA DYSENTERIAE

 
 
Author(s):  AHMADI DANESH HORA, SAADATI MOJTABA*, DOROUDIAN MOHAMMAD, YAKHCHALI BAGHER, KARKHANEH ALI ASGHAR, HOSSEINI SAYED MOSTAFA, ZARE MOKHTAR, HASHEMI MEHRAD
 
* DEPARTMENT OF CELL BIOLOGY, FACULTY OF SCIENCE, IMAM HOSSEIN UNIVERSITY, TEHRAN, IRAN
 
Abstract: 

Background: Shigellosis disease is the major causes of morbidity in children with diarrhea in Iran.
ThevirG (icsA) gene plays a key role in pathogenesis and ability of invasion in shigella. The aim of this study was cloning, sequencingvirG gene and developing a mutant construct pGEM
D 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.
ThevirG 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 ofvirG gene by using enzymatic digestion was removed. Finally, pGEM
D virG construct was transformed toE. 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
D virG construct contains a mutant construct ofvirG gene which 1751 bp was deleted through enzymatic digestion reaction and transformed inE. 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 attenuatedShigella dysenteriae vaccine candidate.

 
Keyword(s): ALLELIC EXCHANGE, SHIGELLA DYSENTERIAE, VIRG GENE, LIVE ATTENUATED VACCINE CANDIDATE
 
References: 
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