Description
This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain h-L972-derived poly(A)+ RNA at the log phase by the Linker-Primer method by Prof. H. Nojima of Research Institute for Microbial Diseases, Osaka University. cDNAs in this library are unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme sites of NotI and BamHI (BglII)-SmaI adaptor. The pLZ3 vector used in this library can replicate both in S. pombe and E. coli and express S. pombe genes in mammalian cells as it contains an SV40 promoter.
Application 1: PCR cloning of known or unknown genes.
Prepare primers for known or unknown genes (cDNAs) and amplify by PCR from this library followed by cloning to an appropriate vector. The cloned cDNAs are useful for identifying the coding region, large-scale protein production, and preparation of probes, etc. Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression levels of mRNA of the gene of interest.)
Application 2: Cloning cDNAs by functional complementation of the corresponding S. pombe mutants.
