|Notice: Suspension of Services Due to the Inspection work of NIG|
For the inspection work of the institute, services on our website will be stopped during the period of time stated below. We apologize for the inconvenience and thank you for your understanding in this matter.
Fri. 12th Nov. 2021 16:00 - Sun. 14th Nov. 13:00 (JST)
Gene Disrupted Strains - About
PCR experiments for 2514 strains were done as following. As first inspection, two parts of each genome were checked for all strains. One part was the region between initiation codon of the putative disrupted gene and pMUTIN vector sequence(1). The other part was the region between termination codon of the putative disrupted gene and pMUTIN vector sequence(2). Then the strains were confirmed as gene disrupted strains, in which an expected DNA fragment was amplified. Next, as second inspection, the region between initiation codon and termination codon of the putative disrupted gene was checked for the strains in which neither PCR was amplified in the first inspection(3). Then the strains were also confirmed as gene disrupted strains, in which an estimated 10kb DNA fragment including the disrupted gene and pMUTIN vector sequence was amplified.
From above two inspections, a wild type strain and 2090 gene disrupted strains are available for the distribution. (Keita Aoki)
During cultivation of the gene disrupted mutants, erythromycin should be added to culture at final concentrations of 0.5 ug/ml to prevent pMUTIN from popping-out. Although rate of popping-out may depend on individual mutants, the rate in strain MGNA-B928, for example, is about 25%.
Quality verification of gene disrupted strains was largely performed. As a result, 243 strains have passed the test for distribution. 2,335 strains including a wild-type strain are now ready to be distributed (April, 2016).