A robust method for the in vivo cloning of large gene clusters was developed based on homologous recombination (HR), requiring only the transformation of PCR products into Escherichia coli cells harboring a receiver plasmid. Positive clones were selected by an acquired antibiotic resistance, which was activated by the recruitment of a short ribosome-binding site plus start codon sequence from t...

We report the map-based cloning of the leaf rust resistance gene Lr21, previously mapped to a gene-rich region at the distal end of chromosome arm 1DS of bread wheat (Triticum aestivum L.). Molecular cloning of Lr21 was facilitated by diploid/polyploid shuttle mapping strategy. Cloning of Lr21 was confirmed by genetic transformation and by a stably inherited resistance phenotype in transgenic p...

Stem cell (SC) therapy is one of the most promising methods clinical medicine. Although stem cell-containing products are still being investigated in trials, some them already approved for treatment many countries. Therefore, modern medicine providing basic understanding SC subtypes, their properties and potential risks should be incorporated educational programs medical universities.
 The...

Targeted mutagenesis is one of the major tools for determining the function of a given gene and its involvement in bacterial pathogenesis. In mycobacteria, gene deletion is often accomplished by using allelic exchange techniques that commonly utilise a suicide delivery vector. We have adapted a widely-used suicide delivery vector (p1NIL) for cloning two flanking regions of a gene using ligation...

Cloning, mutagenesis and complementation of virulence factors are key steps to understand the mechanisms of bacterial pathogenesis and cloning vectors are routinely utilized for these processes. We have investigated the effect of the presence of commonly used cloning vectors on the survival of the intracellular bacterial pathogen Salmonella during macrophage infection. We demonstrate that the p...

In the past two years, considerable progress has been made in the mapping and cloning of human deafness genes. Highlights are the chromosomal localization of at least five genes for autosomal forms of non-syndromic deafness and, more recently, the cloning of an X-linked deafness gene, DFN3, and the Usher syndrome type IB gene. This last gene encodes a myosin-like protein and was identified as t...

The present study was undertaken to clone, express rabies virus glycoprotein (RVG) and to identify potential T-cell epitopes on it. RVG gene (1590 bp) was amplified using gene specific primers. The amplified product was cloned into pTZ57R/T cloning vector by TA cloning. RVG gene was subcloned into pcDNA3.1 (+) expression vector. In this study, cloning and expression of rabies virus glycoprotein...