A novel yeast three-hybrid (Y3H) vector pBT was developed, which contains a tetracycline (Tet)-sensitive transactivator (tTA) expression unit and a Tet-responsive element (TRE)-driven 3rd protein expression unit within a single plasmid. To optimize tTA expression levels, several promoters for driving tTA expression were tested, and the weakest human cytomegalovirus (CMV) promoter showed the bes...

Blinatumomab, the bispecific T cell engager, has been demonstrated as the most successful BsAb to date. Throughout the past decade, vector design has great importance for the expression of monoclonal antibody in Chinese hamster ovary (CHO) cells. It has been indicated that expression plasmids based on the elongation factor-1 alpha (EF-1 alpha) gene and DHFR selection marker can be highly effect...

Saccharomyces cerevisiae was engineered to express different amount of heavy (H)- and light (L)-chain subunits of human ferritin by using a low-copy integrative vector (YIp) and a high-copy episomal vector (YEp). In addition to pep4::HIS3 allele, the expression host strain was bred to have the selection markers leu2(-) and ura3(-) for YIplac128 and YEp352, respectively. The heterologous express...

Background and Aims: The aim of this study was cloning and expression of rabies virus glycoprotein by a eukaryotic expression plasmid pcDNA3.1(+) in BSR cell line. This construct might be used for a potential DNA vaccine. Materials and Methods: Glycoprotein gene was synthesized and cloned into pBluescript vector and then sub cloned into eukaryotic expression vector (pcDNA3.1(+)). After verifica...

Blinatumomab, the bispecific T cell engager, has been demonstrated as the most successful BsAb to date. Throughout the past decade, vector design has great importance for the expression of monoclonal antibody in Chinese hamster ovary (CHO) cells. It has been indicated that expression plasmids based on the elongation factor-1 alpha (EF-1 alpha) gene and DHFR selection marker can be highly effect...

A flocculating yeast Saccharomyces cerevisiae ura3 was transformed by the method based on treatment of intact cells with lithium acetate plus single-stranded carrier DNA using the shuttle vector pYAC4. The transformation efficiency was above 10(3) transformants per microgram of plasmid DNA which is similar to other described yeast transformation systems. Under selective pressure, the transforme...

Functional complementation of the Saccharomyces cerevisiae glycogen branching enzyme deficiency was screened to isolate human cDNAs that encode this enzyme. Human hepatoma cell line HepG2-derived cDNA libraries using the pAB23BXN yeast expression vector yielded four cDNAs capable of complementing the glc3::TRP1 glycogen branching enzyme mutation. Complementation was recognized by an altered iod...

The ratio between proteins P27 and replicase of Cocksfoot mottle virus (CfMV) is regulated via a -1 programmed ribosomal frameshift (-1 PRF). A minimal frameshift signal with a slippery U UUA AAC heptamer and a downstream stem-loop structure was inserted into a dual reporter vector and directed -1 PRF with an efficiency of 14.4 +/- 1.9% in yeast and 2.4 +/- 0.7% in bacteria. P27-encoding CfMV s...