DEVELOPMENT OF DOUBLE GENE MAMMALIAN EXPRESSION VECTOR

Abstract

Recombinant vectors are valuable tools in the biopharmaceutical industry with a number of novel vectors being developed every day. These Antibodies are often expressed in mammalian cells by cotransfecting light chain and heavy chain containing plasmids. However, co-transfection can lead to variable in the copy number of both heavy chain and light chain, there by affecting the protein productivity. Double gene expression vectors can overcome these problems. In the current design The first transcriptional unit comprises sequences for the CMV promoter, multiple cloning site (MCS) and a Polyadenylation Sequence and the second transcriptional unit comprises sequences for the SV40 promoter, MCS and polyadenylation sequence. The double gene expression vector (pUB-C-S) was constructed by ligating the BglII and BamHI fragment (974bp) form pSI plasmid with BglII digested pCI plasmid. The presence of two independent transcriptional units in the recombinant plasmid was confirmed by colony PCR.