Trace the events leading to development of M13mp8 from M-13 phage.( 6 marks ) .NOTE: Please...

Trace the events leading to development of M13mp8 from M-13 phage.( 6 marks )

.NOTE: Please answer sensibly !

Expert Solution

Ans M13 is a single stranded Dna bacteriophage that infect male Escherichia coli strains. A chronic infection is set up during intracellular replicative form of phage (RF-1, double stranded Dna) serve as template for single stranded Dna prodution.

M13mp8 is M13 derivative. It contain multiple cloning site(MCS) within a portion of the gene for beta- galactosidase. This Dna segment correct a defect in beta- galactosidase production by the host strain. TheMCS region of M13mp8 contain 9 or 13 unique restriction endonuclease site.

Foreign Dna incorporated at MCS in loss of beta - galactosidase production and the inability to use lactose as a carbon source . Such lactose negative strain are detected as white plaqueson medium containing X- Gal , a chromogenic substrate for beta - galctogalactosidase.

The interacellular ,double stranded (RF-1) form of M13 phage can be manipulated like plasmid DNA ,while extracellular, single stranded Dna is an excellent source of material for determining the sequence of insert dna by the dideoxy procedure . The multiple cloning site of M13mp9/19 is in the opposite orientation relative to the cloning site of M13mp8/18.

gladiator answered 2 years ago

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