This is a service for the generation of BAC-based transgene vectors for expressing reporter genes like LacZ or GFP, or Cre or other functional gene products.  This service assumes one recombineering step and typically takes 3 - 6 weeks.  A typical use of this approach is to insert a cassette encoding a reporter gene or other functional protein into a BAC containing a gene of interest, such that the cassette will be expressed under control of the gene’s regulatory elements.   An FRT-flanked antibiotic marker is used to facilitate selection of the insertion, and the marker can be easily removed by FLP expression in bacteria.  This service can also usually be used for investigators who simply want to delete pieces of unwanted DNA from a BAC clone.

Experimental flow for BAC recombineering services: 

1. The principal investigator (PI) schedules a time to meet with Dr. Mortlock (mortlock@chgr.mc.vanderbilt.edu).  Dr. Mortlock is available to design the desired vector or provide consultation, depending on the needs of the PI.  Together they will determine which, if any, reagents must be provided by the PI before vector construction (e.g. specialized expression cassettes) and which reagents will be obtained by Dr. Mortlock on behalf of the PI (e.g. standard, publicly available BAC clones).   The outcome of the meeting will be to create a written plan for vector construction and the final vector design. 
2. Dr. Mortlock creates maps and Genbank files of the vector to be generated.
3. The PI submits the appropriate service form to the TMESCSR including the required documentation, as described on the submission form.
4. The BAC service will be scheduled by the TMESCSR.  Dr. Mortlock will notify the PI of the start date.
5. The PI will be billed the month that service begins.
6. When completed, Dr. Mortlock will provide the PI with the final vector as a glycerol stock of the bacteria stock (whether BAC or plasmid).
7. Intermediate BAC clones (generated as intermediate steps in vector construction) will be stored by the Core as bacterial glycerol stocks. Duplicates of these may be provided to the PI upon request.

Please note that a set of guidelines for BAC recombineering of transgenes and targeting vectors can be found at vanderbiltresearch.org in the TMESCSR protocol collection under "BAC recombineering and transgenesis."