Exotoxin a is one of the toxins which produce by group a streptococcuses. this toxin is a protein soluble substance which causes scarlet fever and different kinds of infections as a super antigen. this toxin has antigenic property and special anti-toxins can neutralize it. in this research, was studied production of streptococcus pyogenes exotoxin a recombinant protein in e.coli by cloning.
Methods
At the first, genome of the bacteria was extracted and used pcr. then agarose gel electrophoresis was performed for detection of size and quality of the resulting amplicons. according to ta cloning kit protocol, the target gene was inserted in to the ptg19 cloning vector and the recombinant plasmid was inserted in to the e.coli xl1blue bacteria. eventually checking the sequencing data obtained from cloned gene in the vectore compared with the refrenece sequenece obtained from the ncbi database, confirm the success of the gene cloning and its correct sequence.
Results
The results showed that pcr product is a fragment with a length of about 708 bais pairs. this is for the first time that the exotoxin a gene was cloned by the ta cloning in the ptg19 cloning vector and the e.coli xl1blue host cell. the results of sequencing was confirmed presence of target gene.
Conclusion
The spea gene was amplified and the recombinant protein production was confirmed by ta cloning with ptg19 cloning vector and e.coli xl1blue host cell.
