knowing the nucleotide sequence of the cholera toxin operon, we designed oligonucleotide primers for its-pcr amplification from local clinical isolates of v. cholerae. the resulting amplification product was cloned in a common puc18 vector. subsequently, a part of this operon encoding the cholera toxin bsubunit (ctb) was reamplified and cloned between the bamh1 and ecor1 sites of the same vecto...

The expression of E74-like factor 5 (ELF5) in epithelial ovarian carcinoma tissues and its effects on biological behavior in ovarian carcinoma cells were assessed in search for a new approach for gene treatment of epithelial ovarian carcinoma. RT-PCR technology was applied to detect the expression of ELF5 mRNA in epithelial ovarian carcinoma (n=49), borderline ovarian epithelial tumor (n=19), b...

A self-regulated high-copy number plasmid containing chloramphenicol resistant gene, for the production of recombinant proteins under the regulation of bacteriophage ?pL promoter, was constructed. The designed 5024 base pair expression plasmid contained a heat sensitive repressor cI857 coding gene to regulate the function of ?pL promoter under heat shock induction. Using the constructed vector,...

  Genetic typing methods of T. gondii strains have been extensively perfected in recent years. From a technical point of view, many tools usable for genetic studied on single-copy loci have been used: RFLP, PCR-RFLP, sequencing, RAPD-PCR and isoenzyme analysis. We described the cloning and sequence analysis of the gene which encodes the major surface antigen (SAG1 or P30) of T. gondii. SAG1 is ...

background and objectives: toxoplasma gondii is an obligatory intracellular parasite which causes severe diseases in the fetus of pregnant women and immunocopmromised patients. serological tests based on recombinant protein are one of the main diagnosis methods for the detection of specific antibodies in serum samples. dense granule antigenic proteins derived from t. gondii (tggras) are potenti...

purpose: in order to express human granulocyte-macrophage colony stimulating factor (hgm-csf) under heat shock. materials and methods: two expression plasmids were constructed based on pbc(sk) plasmid. the expression cassettes in the two plasmids are equipped with a 75 base pair fragment, derived from the pl promoter of the bacteriophage lambda (λ). the plasmids also contain a temperature muta...

background: serological assay based on dense granular (gra) proteins of toxoplasma gondii (t. gondii) is actually the most popular laboratory diagnostic tool to detection of toxoplasmosis. we aimed to construct a recombinant gra7-ptz57rt plasmid vectors that it is suitable for sub-cloning and gra7 protein production. m aterials and methods: souris mice were used for maintaining of t. gondii tac...

The genome of HB V virus of serotype ayw cloned in pBR322 and expression shuttle vector p YES2 were used for construction of the HBsAg chimeric genes and their expression in Saccharomyces cerevisiae. Two recombinant plasmids were constructed. One of them contained the coding sequences for the major polypeptide of surface antigen. Another construct carried the major polypeptide with the pre-S2 a...

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. The expression of heterologous proteins in P.pastoris is fast, simple and inexpensive. In this study, VP2 encoding gene of classical D78 IBDV was amplified using reverse transcription (RT) polymerase chain reaction (PCR) and cloned into pPICZαA vector...

Background: The production of recombinant proteins in Escherichia coli is one of the most valuable achievements in biotechnology, with many therapeutic and diagnostic applications; however, the aggregation and misfolding of proteins that result in the formation of insoluble inclusion bodies is a disruptive factor in this process. Various solubilization and refolding methods can be used to impro...