Construction of pRMES and pTMES plasmids to test their expression ability of Nanobodies via the efficient T7 promoter

Nanobody technology is considered as a promising molecular biology technique performed by means of the genetic engineering of special type of antibodies, existing exclusively in Camelidea. It enables the obtaining of small proteins, referred to as Nanobodies, which are characterized by high stability and solubility, are able to link to their specific antigens. After production, the Nanobody genes are cloned within plasmids of protein expression in bacteria, allowing their stable and continuous production for research and applied purposes. This work aimed to design new plasmids for Nanobody genes cloning in order to ensure a strong expression via the efficient T7 promoter, thus enhancing the quantity of the produced Nanobodies. These plasmids were called pRMES and pTMES and their ability to express Nanobodies, NbBruc02 and Nb16M, was tested. The plasmid pTMES showed an enhanced production condition of this Nanobody. These new plasmids, by their variable characteristics, could represent efficient tools for general production of recombinant proteins, including Nanobodies (author).