CONSTRUCTING A NOVEL PLASMID TO INCREASE PURITY EFFECTIVENESS FOR RECOMBINANT PROTEINS EXPRESSED IN ESCHERICHIA COLI CELLS

Abstract

Nowadays, requirement of supply for recombinant proteins has increased in several fields such as food technology, medical pharmacy, clinical diagnose or environment treatment. The recombinant proteins have become the commercialized products and of that yielded with increasing in a large number per year. Besides, supposing that on extending of the his-tag of the pET111a plasmid may be facilitate for removing his-tag and more effective in protein purification. In this study, nine nucleotides (GCGGCGGCG) coding three alanine residues were added to positions followed hexa-histidine tag (his-tag) on a pET11a plasmid construction. The SDS-PAGE result of each recombinant protein contained the long-modified tag after purification process almost only exhibited single band on gel. Based on alike observed consequences for three recombinant proteins already refined the purity effectiveness reach to upper 98% in the total of existing proteins inside the solution. Hence, the novel pET11a plasmid construction could become an effective plasmid for the aim of harvesting high-purified recombinant proteins.