there are two different co-expression systems including bicistronic; dual-vector or two-promoter to express two different genes simultaneously and also to study protein-protein interactions. bicistronic system has disadvantages e.g. compared with two-promoter system. in this paper, a simple method based on spliced overlap extension by polymerase chain reaction (soe-pcr) technique was demonstrat...

background: follicle stimulating hormone (fsh) plays an essential role in reproductive physiology and follicular development. objective: a new variant of the equine fsh (efsh) gene was cloned, sequenced, and expressed in pichia pastoris (p. pastoris) gs115 yeast expression system. materials and methods: the full-length cdnas of the efshα and efshβ chains were amplified by reverse transcription ...

objectives: in present research we evaluate the expression of this critical enzyme in a eukaryotic system for future use in cheese industry. materials and methods: we have cloned bovine prochymosin gene in methylotrophic yeast, p. pastoris, using ppic9k as an expression vector. the recombinant plasmid was transformed into the host by electroporation, and it was expressed in optimum con­ditions...

Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes ...

background: plants are among promising and suitable platform systems for production of recombinant biopharmaceutical proteins due to several features such as safety, no need for fermentation, inexpensive investment, and fast and easy scale-up. human insulin is one of the most widely used medicines in the world. up to now different expression systems including escherichia coli, yeast and cho hav...

Ubiquitination is a posttranslational protein modification that regulates most aspects of cellular life. The sheer number of ubiquitination enzymes that are present in a mammalian cell, over 700 in total, has thus far hampered the analysis of distinct protein ubiquitination cascades in a cellular context. To overcome this complexity we have developed a versatile vector system that allows the re...

BACKGROUND/AIM Helicobacter pylori is a major health problem. One of the therapeutic approaches is administration of antibody against H. pylori. The methylotrophic Pichia pastoris is a suitable host for expression of recombinant antibody fragments. The aims of this study were the expression and the evaluation of camelid nanobody in the yeast Pichia pastoris. MATERIALS AND METHODS The camelid-...

A new inducible yeast expression vector, pXS7, was constructed by using the promoter and terminator sequences from the Saccharomyces cerevisiae SOR1 gene, which codes for the sorbitol dehydrogenase protein. We cloned the coding sequence of the Saccharomyces YEF3 gene in this vector and demonstrated an increase in YEF3 protein levels when cells were grown in the presence of the sugar sorbitol.

A method for linking any standard yeast artificial chromosomes (YAC) is described. YACs are introduced into the same cell and joined by mitotic recombination between the vector arms and the homologous sequence in a linking vector; several YACs can be recombined sequentially. The linking vectors also contain the beta-galactosidase gene as an expression reporter in mammalian cells.

The methylotrophic yeast Pichia pastoris has been successfully used for the expression of many heterologous proteins. The level of expression of some of these proteins depends on the copy number of the gene inserted into the yeast genome. Several methods have been reported in the past few years for the isolation of multicopy transformants. One of these methods used an expression vector that con...