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The 'helper' phage infects the bacterial host by first attaching to the host cell's pilus and then, after attachment, transporting the phage genome into the cytoplasm of the host cell. Inside the cell, the phage genome triggers production of single stranded phagemid DNA in the cytoplasm. This phagemid DNA is then packaged into phage particles.
The first recombinant DNA molecule was created by Paul Berg in 1972 when he combined DNA from the monkey virus SV40 with that of the lambda phage. [ 12 ] Experiments on hereditary material during the time of the Hershey–Chase experiment often used bacteriophages as a model organism .
The structures of the phage capsid and of some other phage proteins are available from the Protein Data Bank. [6] The single-stranded Ff phage DNA runs down the central core of the phage, and is protected by a cylindrical protein coat built from thousands of identical α-helical major coat protein subunits coded by phage gene 8.
Like the two-hybrid system, phage display is used for the high-throughput screening of protein interactions.In the case of M13 filamentous phage display, the DNA encoding the protein or peptide of interest is ligated into the pIII or pVIII gene, encoding either the minor or major coat protein, respectively.
Electroporation allows cellular introduction of large highly charged molecules such as DNA which would never passively diffuse across the hydrophobic bilayer core. [2] This phenomenon indicates that the mechanism is the creation of nm-scale water-filled holes in the membrane. [ 42 ]
The "early", "middle" (DNA replication), and "late" genes (virus structure), roughly represent the time course of gene expression. [76] Bacteriophage genomes can be highly mosaic, i.e. the genome of many phage species appear to be composed of numerous individual modules. These modules may be found in other phage species in different arrangements.
Assembled major coat protein, exploded view. The virion is a flexible filament (worm-like chain) about 6 nm in diameter and 900 nm long.Several thousand copies of a small (50 amino-acid residues) elongated alpha-helical major coat protein subunit (the product of gene 8, or p8) in an overlapping shingle-like array form a hollow cylinder enclosing the circular single-stranded DNA genome.
Nucleofection is an electroporation-based transfection method which enables transfer of nucleic acids such as DNA and RNA into cells by applying a specific voltage and reagents. Nucleofection, also referred to as nucleofector technology , was invented by the biotechnology company Amaxa.