CYP3A4 probably has the broadest catalytic activity of any cytochrome P450. It is a crucial task to test new drug candidates in a reliable system for their ability to induce expression of this enzyme. Firstly, a total of 300 bp core distal enhancer of CYP3A4 XREM region (-7972/-7673) were amplified from human genomic DNA. The PCR product was then ligated into a human secretory alkaline phosphat...

conclusions: e. coli herb-resistance plasmid can replicate and be expressed in s. aureus. results: the identified 45 kb herb-resistance plasmid could be transferred from e. coli cp9 isolates to e. coli dh5α. as a consequence e. coli dh5α transconjugant mic increased from 0.0125 g/ml to 0.25 g/ml. the plasmid was easily transferred from e. coli cp9 strain to s. aureus rn450rf with a mean transfe...

A whole cell extract prepared from Schizosaccharomyces pombe was shown to be active in an assay for repair of plasmid DNA damaged by either ultraviolet (UV) light or gamma-radiation. The assay allows for analysis of repair synthesis at single-strand nicks generated by gamma-rays and analysis of the incision step and repair synthesis in UV-light-damaged DNA. Repair synthesis of DNA damaged by ei...

Reporter gene assays are important tools for evaluating gene expression. A frequently used assay measures the activity of β-galactosidase (β-gal) expressed from lacZ in plasmid or genomic constructions. Such constructions are often used to interrogate the ability of DNA (query DNA), potentially encoding a transcription factor, to regulate in trans the expression of a promoter fused to the repor...

Plasmid DNA is being used successfully as a gene delivery vector in a variety of clinical applications. Similar to other pharmaceutical products for clinical use, the plasmid vectors must meet rigorous purity standards. One important contaminant is the DNA of the host cell used to produce the plasmids. We have developed a new method to accurately quantitate E. coli host-cell DNA in plasmid prep...

A significantly improved DNA polymerase fidelity assay, based on a gapped plasmid containing the lacZα reporter gene in a single-stranded region, is described. Nicking at two sites flanking lacZα, and removing the excised strand by thermocycling in the presence of complementary competitor DNA, is used to generate the gap. Simple methods are presented for preparing the single-stranded competitor...

Molecular detection of herpes simplex virus (HSV) DNA is recognized as the reference standard assay method for the sensitive and specific diagnosis of central nervous system infections caused by HSV. In this study, a molecular assay based on real-time PCR on the LightCycler (LC) instrument was evaluated and compared with a home-brew molecular assay. The detection limit of the LC assay was deter...

background: echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus echinococcus . iran is known as endemic region for this infection in the world. vaccination has been considered as a good prevention method for this disease. recombinant vaccines containing eg95 protein, against e. granulosus , has shown a high d...

BACKGROUND Avian hepatitis E virus (HEV) is the main causative agent of big liver and spleen disease, as well as hepatitis-splenomegaly syndrome in chickens. To date, conventional reverse transcriptase polymerase chain reaction (RT-PCR) and nested RT-PCR methods have been used for the diagnosis of avian HEV infection in chickens. However, these assays are time consuming, inconvenient, and canno...

We previously demonstrated that plasmid-deficient Chlamydia muridarum retains the ability to infect the murine genital tract but does not elicit oviduct pathology because it fails to activate Toll-like receptor 2 (TLR2). We derived a plasmid-cured derivative of the human genital isolate Chlamydia trachomatis D/UW-3/Cx, strain CTD153, which also fails to activate TLR2, indicating this virulence ...