The present invention relates to the construction and insertion of a wide spectrum vector for Gram-negative bacteria, having a gene sequence which, when expressed, allows a mercury ion chelating protein to be anchored onto the cell surface of gram-negative bacteria. For that purpose, the merR regulatory gene was isolated from the pMOL30 plasmid of Cupriavidus metallidurans, line CH34 (SEQ ID NO. 1) and inserted into the pGEMT cloning vector, producing the PGEMT-Hg plasmid (SEQ ID NO. 2). The expression vector containing the sequence corresponding to the cassette for expressing and anchoring heterologous proteins in Gram-negative bacteria under the control of the pan promoter (SEQ ID NO. 3) derived from the pCM2 plasmid was obtained by polymerase chain reaction (PCT). The merR gene was merged with the amplified expression vector, leading to the construction of the pCMHg plasmid (SEQ ID NO. 4). In addition, the present application provides mutant strains of Gram-negative bacteria containing said recombinant plasmid, a method of obtaining same, and discloses the possible use of the transgenic strain for adsorbing mercury ions in environmental bioremediation processes.
PATENT NUMBER
WO2012145814A2
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