| 主权项 |
1. A method, comprising:
introducing into yeast cells (a) copies of a linearized yeast artificial chromosome (YAC) having at each end a sequence of at least 20 contiguous nucleotides, and (b) a set of linear bacteriophage genomic fragments, each genomic fragment comprising at each end a sequence of at least 20 contiguous nucleotides, wherein the set of genomic fragments, when recombined via homologous recombination of the end sequences, forms a nucleic acid that encodes a viable bacteriophage, wherein the set comprises
(i) a first subset of linear bacteriophage genomic fragments that together encode proteins that form a capsid head of one type of bacteriophage selected from T3 bacteriophage, T4 bacteriophage, T5 bacteriophage, T7 bacteriophage, K1F bacteriophage, K11 bacteriophage, YppR bacteriophage and SP6 bacteriophage, and(ii) a second subset of linear bacteriophage genomic fragments that together encode proteins that form a tail of another type of bacteriophage selected from T3 bacteriophage, T4 bacteriophage, T5 bacteriophage, T7 bacteriophage, K1F bacteriophage, K11 bacteriophage, YppR bacteriophage and SP6 bacteriophage that is different from the type of bacteriophage of (i), wherein one of the two end sequences of each bacteriophage genomic fragment of (i) and (ii) is homologous to only one other end sequence of an adjacent genomic fragment or to an end sequence of a linearized YAC of (a); and culturing the yeast cells to permit homologous recombination of the end sequences of the linear bacteriophage genomic fragments of (b) and the end sequences of a linearized YAC of (a), thereby producing a recombined YAC::phage construct that encodes a viable recombinant bacteriophage having the capsid head of one type of bacteriophage and the tail of another type of bacteriophage. |
