Wild-type T7 is a superb organism for discovering the primary components of a natural biological system. However, our experience indicates that the original T7 isolate is not best suited for understanding how all the parts of the phage are arganized to encode a functioning whole. We decided to engineer a surrogate genome, which we designated T7.1, that would be easier to study and extend.
Goals
We wanted to insulate and enable independent manipulation of all identified genetic elements.
We wanted the T7.1 genome to encode a viable bacteriophage; at the start of this work, we were uncertain how many simultaneous changes the wild-type genome could tolerate.
Reannotation of the wild-type T7 genome, thus defining the functional genetic elements
Specification of T7.1 genome design and sequence
Construct sections individually
Construct chimeric phages that contain replace a single wild-type section with a rebuit section
Combine sections of rebuilt phage into a single rebuilt phage
Characterize chimeric phage
Progress
Reannotation of the T7 Genome -- The wild-type T7 genome is a 39,937 base pair linear double-stranded DNA molecule. We annotated the genome by specifying the boundaries of the following functional genetic elements: 57 open reading frames, 57 putative RBSs encoding 60 proteins, and 51 regulatory elements controlling phage gene expression, DNA replication, and genome packaging. A genbank file of the reannotation can be found here.
