Construction of pcDNA/fimH cassette as a DNA vaccine candidate against urinary tract infection and evaluate its expression in COS7 cell line
Trauma Monthly: 15 (4); 203-206 Article Type: Research Article
M, Imani Fooladi
A A. Construction of pcDNA/fimH cassette as a DNA vaccine candidate against urinary tract infection and evaluate its expression in COS7 cell line,
Online ahead of Print
Aims: The most important cause of urinary tract infection is Uropathogenic Escherichia coli (UPEC) which produces type I pili. UPEC has intracellular duplication after invasion to bladder epithelial cells therefore, cellular immunity is important in this case. This study was performed in order to design a gene cassette inducing cellular immune system against Uropathogenic Escherichia coli.
Materials & Methods: At first, fimH gene was amplified by PCR. PCR product was inserted to pcDNA.1 eukaryotic expression vector and the recombinant vector was confirmed by sequencing. The clone was inserted into COS7 cell and then assessed by RT-PCR in order to evaluate gene expression and the correct function of the gene cassette. Gene expression at mRNA level was evaluated by florescence microscope.
Results: fimH gene was well amplified by PCR. PCR product was inserted to pcDNA.1 eukaryotic expression vector and pcDNA/ fimH gene cassette was constructed. COS7 cells were transfected in order to evaluate the correct function of this cassette. Expression of fimH gene in COS7 was confirmed by RT-PCR.
Conclusion : pcDNA/ fimH cassette could express inserted fimH gene in eukaryotic cells and is a valuable DNA cassette for urinary tract infection vaccination.
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