Vanillyl alcohol oxidase (VAO) from Penicillium simplicissimum is a covalent flavoprotein that has emerged as a promising biocatalyst for the production of aromatic fine chemicals such as vanillin, coniferyl alcohol and enantiopure 1-(4’-hydroxyphenyl) alcohols. The largescale production of this eukaryotic enzyme in Escherichia coli has remained challenging thus far. For that reason an alternat...

RNA interference (RNAi) is one of the primary machineries involved in regulation gene expression using small double-stranded (dsRNA) eukaryotic cells. MicroRNA (miRNA) a class non-coding RNAs, regulating through canonical and non-canonical ways. Previous studies have shown that miRNA coding sequences make up 1% human genome currently 1917 miRNAs are displayed miRBase database. Expression levels...

AIM:To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector cosmid pTM3 and to express HCV core antigen in HepG2 cells.METHODS:Core gene cDNA of HCV was introduced into eukaryotic expression vector cosmid pTM3.Using vaccinia virus/bacteriophage T7 hybrid expression system, HepG2 cells were transfected with the recombinant plasmid pTM3-Q534 by lipofectin.RESU...

Trigger factor and DnaK protect nascent protein chains from misfolding and aggregation in the E. coli cytosol, but how these chaperones affect the mechanism of de novo protein folding is not yet understood. Upon expression under chaperone-depleted conditions, multidomain proteins such as bacterial beta-galactosidase (beta-gal) and eukaryotic luciferase fold by a rapid but inefficient default pa...

Most of the 231 unique membrane protein structures (as of 3/2010) are of bacterial membrane proteins (MPs) expressed in bacteria, or eukaryotic MPs from natural sources. However eukaryotic membrane proteins, especially those with more than three membrane crossings rarely succumb to any suitable expression in bacterial cells. They typically require expression in eukaryotic cells that can provide...

A general method is described for the site-specific genetic encoding of cyanine dyes as non-canonical amino acids (Cy-ncAAs) into proteins. The approach relies on an improved technique for nonsense suppression with in vitro misacylated orthogonal tRNA. The data show that Cy-ncAAs (based on Cy3 and Cy5) are tolerated by the eukaryotic ribosome in cell-free and whole-cell environments and can be ...

The expression vectors are designed for expression and purification of normal or recombinant gene of interest. There is a wide variety of stable and transferable selectable mammalian expression vectors.The vector pCDM8 and its derivatives, pcDNAI/Ampicilline are widely used for cloning and analyzing of genes in higher eukaryotic cells. The vectors pRC/CMV, pcDNA3 and pRC/RSV are designed for hi...

The efficacious delivery of eukaryotic expression plasmids to inductive cells of the immune system constitutes a key prerequisite for the generation of effective DNA vaccines. Here, we have explored the use of bacteria as vehicles to orally deliver expression plasmids. Attenuated Salmonella typhimurium aroA harbouring eukaryotic expression plasmids that encoded virulence factors of Listeria mon...

background permanent antigenic variation of influenza viruses causes a major concern to develop an effective human influenza vaccine. conserved antigens are new vaccine candidates because it is not necessary to match the prepared vaccine with circulating strains. ion channel m2 protein is conserved among all influenza a viruses, allowing the virus to enter host cells. objectives to prepare an e...

epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant dna techniques. the aim of this study was to sub-clone the peroxisomal protein (pep) cdna into a mammalian expression vector leading to the formation of a  chimeric pep-cdna containing the flag epitope. the flag-pep recombinant cdna was construc...