Gene Expression ServiceServices

• Subcloning of genomic fragments from bacterial or P1-derived artificial chromosomes (BACs/PACs) into high copy plasmids (up to 15kb)
• Subcloning fragments from plasmid to plasmid (up to 15kb)
• Trimming BAC clones (BAC-shaving): deletion of unwanted fragments in BAC clones
• Deletion of sequences from 1bp (counter-selection system) up to 50kb (direct replacement or making use of inserted LoxP/FRT/Rox sites)
• Removal of LoxP/FRT/Rox flanked sequences by bacterial Cre/Flp/Dre expression
• Insertion of functional cassettes (e.g. LacZ, Cre, eGFP, NLS, any given short protein coding tag, coupled cassettes using IRES or P2A/T2A elements, any custom made cassette) into any chosen position in a BAC or plasmid backbone
• Insertion of point mutations at any chosen position
• Design and generation of custom expression cassettes

All steps must involve selectable markers to screen for successful recombination events. Selected applications and services include: (A) Sub-cloning sequences from BACs/PACs into high copy plasmids. (B) Deletion of sequences (from 1bp up to 50kb, with or without leaving selectable marker genes). (C) Insertion of functional cassettes into any chosen position. (D) Generation of complex BAC transgenic constructs (e.g. reporter lines, Cre/Flip-drivers). (E) Point mutations at any chosen position in either BAC or plasmid backbones. (F) Design and generation of custom tissue-specific expression cassettes (e.g. inducible transgenes, defined promoter-containing transgenes). (G) Assembly of knock-in constructs. We provide services for both, generating constitutive or conditional targeting constructs