Construction of a Nisin-Controlled Expression Vector, a Derivative of pMSP3535 for Alternative Selectable Marker
Year: 2009
Type: article
Abstract: The plasmid pMPS3535 is a commonly-used nisin-inducible expression vector for homologous and heterologous genes in a variety of gram-positive bacteria.However, a number of limitations associate with this plasmid, including the use of the erythromycin resistance (Em r ) marker.Em r is somewhat ubiquitous element in shuttle vectors, bacterial plasmids, and even in bacterial chromosomes, which limited its application in hosts naturally resistant to erythromycin and in combination with Em r -carrying vectors.To overcome this limitation, the Em r marker was displaced by an alternative tetracycline resistance maker encoded by tetL gene.The resulting vector, termed pMSP3535TLH, has been proved to be able to transform Enterococcus faecalis OG1RF, Lactococcus lactis 2301, and Escherichia coli DH5α to tetracycline resistance whose transformants were readily selected on selective plates supplemented with tetracycline.By taking advantage of the Em r marker removal to simultaneously excise a segment of copF gene, a negative regulator of copy number, the constructed vector has potential to become a high copy number derivative of the plasmid pMSP3535.
Author: Monchaya Rattanaprasert
Institution: The Ohio State University
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2739532859