One of the most powerful strategies to investigate a gene's function is to inactivate, or "knockout", the gene by replacing it or disrupting it with an piece of DNA designed in the lab. Specially …

In a gene targeting experiment, a replacement vector directs the disruption of an essential coding exon/s of a gene through the insertion of a positive drug resistance marker. This replacement …

2008年5月29日 · We describe here a streamlined procedure for targeting vector construction, which often is a limiting factor for gene targeting (knockout) technology. This procedure …

Gene replacement: Mutant alleles cloned into the pKO3 gene replacement vector are electroporated into recombination proficient strains (eg. EMG2) and allowed to recover for 1 h …

Deletion of sti on Streptomyces plasmid pIJ101, made its derivative pHZ1358 an efficient vector for gene disruption and replacement. Here, we further optimized pHZ1358 by the construction …

Knockouts are used to define the overall requirement for a gene and to model loss of function mutations. Knockins introduce heterologous coding sequences such as reporters or DNA …

The allelic replacement vector, designated here as pDG1, expresses an X-Gal hydrolyzing enzyme (BgaB) that can be used for blue/white screening allowing identification of colonies …

2017年9月19日 · We have developed a rapid streamlined method to create clean deletion mutants in P. aeruginosa through direct transformation, eliminating the need for the creation of …

2024年12月28日 · This mutated donor DNA vector is co-transformed into the P. sojae knockout line together with Cas9 and two new RNA templates targeting the substitution gene. In this …

2015年10月10日 · We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded …