3 results about "RAD51" patented technology

The present application belongs to the field of microbial metabolic engineering and industrial microorganism technology, and particularly relates to a method for stably integrating exogenous DNA into Pseudozyma roseolata, an iterative gene editing method and application thereof. The preservation number of the strain MYL-2 is CCTCC M 20252968, and the strain can efficiently accumulate oil under nitrogen-limited culture conditions, the main fatty acid components in the cells are C16-C18 fatty acids, and the strain has application potential as a microbial oil production strain. The present application provides a method for stably integrating exogenous DNA into Pseudozyma roseolata, and constructs a recombinant expression vector pNFH-loxp-RAD51 and a Cre recombination vector containing a loxP site, and for the first time, precise removal and repeated use of a screening marker mediated by a Cre-LoxP system are realized in the strain, thereby laying a foundation for multi-round gene editing and efficient engineering modification of the strain.

This invention relates to the field of biomedical technology, disclosing succinylated modified RAD51 protein, formulations for promoting RAD51 succinylation, and their applications. The method includes: seeding and culturing cells and transfecting them with a tagged RAD51 expression plasmid; adding sodium succinate and an HDAC11 inhibitor to the culture medium for continued culturing to allow intracellular modification accumulation; collecting the supernatant after cell lysis and incubating it with magnetic beads containing anti-tag antibodies under cold conditions; washing and eluting with an elution buffer containing the tagged peptide; performing immunoblotting analysis using antibodies that specifically recognize the modified site to obtain the succinylated modified RAD51 protein. This invention, by adding sodium succinate and an HDAC11 inhibitor, provides a substrate and blocks the demodification pathway. Combined with magnetic bead purification and competitive elution, it avoids disrupting the protein conformation, obtaining a protein that retains its native activity and has a high level of modification, providing a core component for screening targeted drugs.

The present invention relates to a CRISPR Cas9 vector and its uses, and more specifically, to a CRISPR Cas9 vector with increased gene editing efficiency. The CRISPR Cas9 vector of the present invention expresses the RAD51 gene, and when the CRISPR Cas9 vector of the present invention is used, the efficiency of gene editing increases by more than 2 times, so genes can be edited with higher efficiency and accuracy.