Recent progress in the development of molecular genetic methods enables the manipulation of genes in intact mammalian organisms. The power of such techniques to elucidate complex biological systems was initially recognized and exploited by developmental biologists and immunologists. More recently, the utility of these approaches for examining neural gene function in the context of the intact or...

The advanced cloning-free PCR-based allele replacement strategy (Erdeniz et al., 1997) has been followed in the present study. The principle of this strategy is graphically depicted in Figure 1a. In the gene targeting experiment performed in the present study, the uracil auxotrophy (Ura-) of CEN.PK 113-5D strain has been exploited. Thus, cells where the gene targeting substrate integrated corre...

Gene targeting by single-stranded oligodeoxyribonucleotides (ssODNs) is emerging as a powerful tool for the introduction of subtle gene modifications in mouse embryonic stem (ES) cells and the generation of mutant mice. Here, we have studied the role of ssODN composition, transcription and replication of the target locus, and DNA repair pathways to gain more insight into the parameters governin...

Gene targeting is a genetic technique to modify an endogenous DNA sequence in its genomic location via homologous recombination (HR) and is useful both for functional analysis and gene therapy applications. HR is inefficient in most organisms and cell types, including mammalian cells, often limiting the effectiveness of gene targeting. Therefore, increasing HR efficiency remains a major challen...

Ends-in and ends-out refer to the two arrangements of donor DNA that can be used for gene targeting. Both have been used for targeted mutagenesis, but require donors of differing design. Ends-out targeting is more frequently used in mice and yeast because it gives a straightforward route to replace or delete a target locus. Although ends-in targeting has been successful in Drosophila, an attemp...

Genus Brachyspira,as Gram negative anaerobic bacteria, colonize in dogs intestine. The aim of the current study was to determine the prevalence of Brachyspira spp. for the first time in Iran and rapid identification of Brachyspira spp. in dogs by a new designment of a species-specific primer set for B. canis. One hundred fifty-one fecal samples were obtained ...

The modification of an endogenous gene into a designed sequence by homologous recombination, termed gene targeting, has broad implications for basic and applied research. Rice (Oryza sativa L.), with a sequenced genome of 389 Mb, is one of the most important crops and a model plant for cereals, and the single-copy gene Waxy on chromosome 6 has been modified with a frequency of 1% per surviving ...