Genetics of bacteriophages
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Genetics of Bacteriophages. 1. Lytic growth of bacterio phages. 2. Morphology of selected bacteriophages. 3. Scaffolded assembly. Assembly of complex phages is assisted by scaffolding proteins. These are removed once assembly is complete. 4. ϕ X174.

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Genetics of bacteriophages

Genetics of Bacteriophages

1


Lytic growth of bacterio phages
Lytic growth of bacterio phages

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Scaffolded assembly. Assembly of complex phages is assisted by scaffolding proteins. These are removed once assembly is complete

4


Single stranded dna bacteriophages

  • ϕ by scaffolding proteins. These are removed once assembly is completeX174

The phage fX174 is an icosahedral phage that contains a circular single-stranded

DNA molecule of 5386 nucleotides. It codes for 11 proteins, each of which has been identified. Adding together the size of all those proteins comes to 2330 amino acids, which would require 6990 nucleotides (3 2330) – substantially more than the total length of the genome

Firstly the genes are very tightly packed – there is very little non-coding

sequence in the genome. In most cases, the end of one gene is directly adjacent to (or slightly overlaps with) the start of the next.

Secondly, one of the proteins (A*) corresponds to the C-terminal region of protein A

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Single-stranded DNA bacteriophages

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Single stranded dna bacteriophages1

  • ϕ by scaffolding proteins. These are removed once assembly is completeX174

Single-stranded DNA bacteriophages

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Replication of single strand bacteriophages

  • After the single-stranded by scaffolding proteins. These are removed once assembly is completeϕX174DNA enters the cell, it is converted into a double-stranded molecule (replicative form, RF) by synthesis of the complementary (or ‘minus’) strand (step A).

  • The minus strand provides a template for the production of further copies of the plus strand (step B).

  • These are in turn converted to the double-stranded replicative form by synthesis of the complementary (minus) strand (step C).

Replication of single-strand bacteriophages

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Single stranded dna bacteriophages2

M13 by scaffolding proteins. These are removed once assembly is complete

M13, represent another type of single-stranded phage. These are ‘male-specific’ phages, since they infect the cell by attaching to the tips of the pili specified by the F plasmid

As the replication cycle proceeds, one of the phage proteins that is produced is able to bind to the single-stranded DNA and divert it into the production of phage particles by targeting it to the cell membrane

where it is extruded from the cell, with phage coat proteins being polymerized around it during its passage through the membrane

Single-stranded DNA bacteriophages

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Single stranded dna bacteriophages3

M13 by scaffolding proteins. These are removed once assembly is complete

Single-stranded DNA bacteriophages

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Rna containing phages

MS2(3600 N) by scaffolding proteins. These are removed once assembly is complete

Another type of male-specific phage is represented by MS2

This is an icosahedral RNA-containing phage which attaches to the sides of the F-pili, rather than to the tip as M13 does.

It is an extremely simple phage, containing some 3600 nucleotides ,coding for just three genes: a coat protein, a maturation protein and a replicase

RNA-containing phages

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Double stranded dna phages

Bacteriophage T4(165Kb) by scaffolding proteins. These are removed once assembly is complete

Double-stranded DNA phages

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Terminal redundancy of bacteriophage T4. Multiple length linear DNA is the substrate for packaging into phage particles. The amount of DNA packaged in longer than the genome size, leading to terminal redundancy (a sequence of about 1600 base pairs at one end of the molecule is repeated in the same orientation at the other end)

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B subtilis phage spo1

  • Control of phage development linear DNA is the substrate for packaging into phage particles. The amount of DNA packaged in longer than the genome size, leading to terminal redundancy (a sequence of about 1600 base pairs at one end of the molecule is repeated in the same orientation at the other end)

B. subtilis phage SPO1

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Replication of bacteriophage lambda DNA. After infection, the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particles

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Cutting dna at the cos sites
Cutting the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particlesλ DNA at the cos sites

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Lysogeny the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particlesStructure of the attachment sites of l. (a) Integration requires site-specific recombination between attP (on the phage) and attB (on the chromosome). (b) After integration, the sites at either end of the prophage (attL and attR) have the same core sequence (shaded) as attP and attB but different flanking sequences

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17 the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particles


18 the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particles


Control of the lytic/ the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particleslysogenic switch in λ. CII stimulates transcription of the cI repressor gene from PE; CIII stabilizes CII. CI represses PL and PR and stimulates PM allowing further synthesis of repressor. Cro represses PL, PR and PM

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19 the cohesive ends are ligated to form a circular molecule which replicates (theta mode) to generate more circular DNA. Later, replication switches to the rolling circle mode, generating multiple length linear DNA for packaging into phage particles


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