Gene identification and DNA construct production

Coordinator:    Prof. Dr. Francis Stewart
Institution: Biotechnologisches Zentrum der Technischen Universität Dresden
The goal of SP1 is to organize the DiGTOP gene list and then generate the corresponding DNA reagents, which include tagged BAC transgenes and targeting constructs.

The gene list has three aspects:

First, genes nominated by the applicants as entry points to areas relevant to disease and therapy.
Second, genes suggested by other NGFN funded programs.
Third, sequential tagging of genes identified as proteomic interactors from the above two categories.

After the initial phase, we anticipate that most work will arise from the third category, with capacity for new suggestions from the first two categories. Sequential tagging to navigate the proteome is central to the proposed research because reciprocal confirmations validate the interactions, as well as being the rational way to map systematically.
The gene list will be updated regularly and will be available on the DiGtoP web site. The updating will include regular interaction with other NGFN funded programs. BAC transgenes and promoterless targeting constructs for C-terminal protein tagging will be generated in a 96-well recombineering pipeline for all chosen genes. These DNA reagents will be supplied to SP4 and SP5 for BAC transgenesis and SP4 for targeting.
A subsidiary goal of SP1 is the development of DNA components and tools to extend the strategy.

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