Question 1

What is the difference between ssDNA and dsDNA bacteriophages?

Accepted Answer

ssDNA bacteriophages contain single-stranded DNA genomes, while dsDNA bacteriophages contain double-stranded DNA. ssDNA phages like ΦX174 have compact genomes with overlapping genes that maximize genetic efficiency. dsDNA phages like T7 and Mu employ more elaborate replication mechanisms and generally have larger genomes, enabling complex interactions with bacterial hosts.

Question 2

How does ΦX174 phage maximize its genetic efficiency?

Accepted Answer

ΦX174 uses overlapping genes where different reading frames encode multiple proteins from the same DNA sequence. This strategy allows the icosahedral ssDNA phage to pack more genetic information into its small genome. The phage replicates via rolling circle replication, forming a double-stranded intermediate before generating new genomes that are released through host lysis.

Question 3

Why does M13 phage exit the host cell differently than ΦX174?

Accepted Answer

M13, a filamentous ssDNA phage, exits through extrusion rather than lysis, allowing continuous phage production while keeping the host alive. This contrasts with ΦX174, which lyses the host to release progeny. M13's sustained production strategy is widely used in molecular biology techniques such as phage display and recombinant DNA technologies.

Question 4

What role does the terminase enzyme play in T7 phage replication?

Accepted Answer

T7 replicates bidirectionally, forming concatemers—long continuous strands containing multiple genome units. The terminase enzyme cleaves these concatemers into individual genome-sized segments for encapsulation into new phage particles. This efficient replication strategy has made T7 a model system in synthetic biology and genetic engineering.

Question 5

How does Mu phage replicate differently from other dsDNA bacteriophages?

Accepted Answer

Mu replicates through transposition rather than conventional rolling circle or bidirectional replication. It integrates into various sites within the bacterial genome and undergoes multiple rounds of transposition before excising itself for packaging into new virions. Mu's transposition duplicates host DNA at insertion sites, often disrupting genes, making it useful for mutagenesis and bacterial genome mapping.

Question 6

What is rolling circle replication in bacteriophages?

Accepted Answer

Rolling circle replication is a mechanism used by ssDNA phages like ΦX174 to generate new viral genomes. The process begins by synthesizing a complementary strand, forming a double-stranded DNA intermediate called the replicative form. This RF serves as a template for continuous synthesis of new viral genomes that are then packaged into virions.

Question 7

How do bacteriophages inject their DNA into bacterial hosts?

Accepted Answer

DNA bacteriophages infect bacteria by injecting their genetic material directly into the host cell. Once inside the bacterial cytoplasm, the phage DNA hijacks the host's cellular machinery for replication. Different phage types then employ distinct replication strategies—such as rolling circle replication in ssDNA phages or bidirectional replication in dsDNA phages—to generate progeny.

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