Transposons have been used in many laboratories worldwide as a powerful tool for insertional mutagenesis to investigate gene function in bacteria (8). The technique has recently been adapted in which a transposon, such as mini-Tn5, delivers a unique identifying sequence of 40 nucleotides (1). This technique, termed signature-tagged mutagenesis (STM), is widely used in the search for virulence-a...

Localized sets of random point mutations generated by PCR amplification can be transferred efficiently to the chromosome of Acinetobacter ADP1 (also known as strain BD413) by natural transformation. The technique does not require cloning of PCR fragments in plasmids: PCR-amplified DNA fragments are internalized by cells and directly incorporated into their genomes by homologous recombination. P...

Background: In reverse genetics studies, minigenomes of negative sense RNA viruses are used to model the process virus replication and evaluate rescue system in course construction recombinant virus. current study, a minigenome (MG) for lentogenic D58 vaccine strain Avian orthoavulavirus 1 (AOAV-1) (Newcastle disease virus) was developed establish genetic this strain. Methods: The constructed b...

A prerequisite for the use of recombinant antibody technologies starting from hybridomas or immune repertoires is the reliable cloning of functional immunoglobulin genes. For this purpose, a standard phage display system was optimized for robustness, vector stability, tight control of scFv-delta geneIII expression, primer usage for PCR amplification of variable region genes, scFv assembly strat...

Background and Aims: Newcastle disease virus (NDV) is one of the major pathogen in poultry. Vaccination is intended to control the disease as an effective solution nevertheless this virus is a growing threat to the poultry industry. F gene open reading frame (ORF) from NDV is 1650 bp, encoding a protein of 553 amino acids that can induce protective immunity alone. The F glycoprotein on the surf...

background: recent researches have demonstrated that human t-cell immunoglobulin mucin 1 (tim-1) glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. therefore,targeting tim-1 could be a potential therapeutic approach against such diseases.objectives: in this study, we aimed to express tim-1 protein ...

To evaluate PCR-generated artifacts (i.e., chimeras, mutations, and heteroduplexes) with the 16S ribosomal DNA (rDNA)-based cloning approach, a model community of four species was constructed from alpha, beta, and gamma subdivisions of the division Proteobacteria as well as gram-positive bacterium, all of which could be distinguished by HhaI restriction digestion patterns. The overall PCR artif...

OBJECTIVE To isolate and construct a cloning vector containing the human papillomavirus (HPV)16-E7 gene as a target for application as a DNA vaccine. METHODS The study was performed in 2005 in Iran. The E7 gene, one of the most important HPV oncoproteins and a target molecule for therapeutic vaccines, was amplified by polymerase chain reaction (PCR). The PCR product was cloned into a suitable...