New multi-purpose high copy number vector with greater mitotic stability for diverse applications in fission yeast Schizosaccharomyces pombe

We have constructed a pUC19-based multipurpose ATG vector in Schizosaccharomyces pombe with higher copy number and mitotic stability possible with commonly used vectors. The vector, having an NdeI site in its polylinker to provide ATG site for expression, carries a greatly truncated version of URA3 gene, URA3m, of Saccharomyces cerevisiae as a selection marker. In addition, it contains the mat2P-right flank region (mat2P-RF) of S. pombe as an autonomous replicating sequence (ARS) and a polylinker with wider choice of restriction sites. While URA3m confers an increase in plasmid copy number up to 200 copies/cell, mat2P-RF imparts greater mitotic stability than the standard ars1 element of S. pombe. Finally, the vector also includes the transcription termination signal of the nmt1 gene (Tnmt1). This basic vector should serve as a versatile tool for studies of gene function in S. pombe.

► A vector has been developed that exists at over 200 copies/cell in Schizosaccharomyces pombe. ► Maintenance of high copy number does not require expensive drugs like geneticin. ► This vector displays high mitotic stability. ► The vector has diverse applications like genomic cDNA libraries, expression libraries, etc. ► It has great potential as an expression vector for protein expression in S. pombe.