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| Cloning of Exotoxin A Gene from Pseudomonas Aeruginosa ATCC27853 Strain and Its Expression in HEK 293T Cells |
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Abstract The full-length coding region of PEA gene was amplified by PCR method from genomic DNA of ATCC27853 Strain of Pseudomonas aeruginosa . The PEA gene was inserted into eucaryotic expression vector pcDNA3.1A to constructe plasmid expression vector pcDNA3.1/PEA . The recombinant plasmids were transfected into HEK 293T cells to be expressed by calcium phosphate mediation. The results showed that cloned PEA gene shared 99% alignment with standard strain PA103. The PEA gene product was detected in the medium with Western blot, which indicated that the PEA gene with original signal peptide could be expressed with secretary form in eucaryotic cells. These lay the foundation for the further research of PEA-based immunotoxin , vaccine adjuvant and vaccine vector.
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Received: 10 March 2008
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