![]() We also generated specific, simultaneous deletions using combinations of spacer variants and inverted repeat variants. Employing these methods, we produced serial modifications encompassing four independent changes which generated a mouse HoxB BAC with fluorescent reporter proteins inserted into four adjacent Hox genes. We show that Lox spacer mutants are very specific, and inverted repeat variants reduce Lox reaction rates through reducing the affinity of Cre for the site, while retaining some functionality. In this work, we investigate the applicability of Lox variant sites for serial and/or simultaneous manipulations of BACs. Techniques such as recombineering and Cre/Lox methodologies have thus become heavily relied upon for such purposes. The size of these constructs, however, prevents the use of conventional molecular biology techniques for modification and manipulation. Bacterial Artificial Chromosomes (BACs) are vital tools in mouse genomic analyses because of their ability to propagate large inserts.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |