User:Slokwong/MODULE 1

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Genome Engineering

Notes on M13 Renovation

M13 genome

  1. Overlapping regions:
    • g2 and g10 (496-831)
    • g9 and g8 (1301-1304)
    • g7 and g9 (1206-1209)
    • g1 and g11 (3916-4242) – g11 is contained in g1, share STOP codon
    • g1, g4, and g11 (4220-4242)
  2. Independent genes:
    • g3, g5, g6

Restriction enzymes used

  • Pst1 digest

M13K07 :

Gene Function Re-engineering Idea
I assembly Modify so that p1 can interact with multiple p4. Then analyze how phage secretion changes. Changing p1-p4 interaction affects how effective the channels are.
II replication Modify such that it nicks the DNA in a way that helps p5 sequester the + strands more effectively, thus controlling the formation of double stranded DNA and perhaps making p5 work more efficiently by reducing competition. Be careful in modifying because it is linked to g10.
III phage tail protein (5 copies) Add myc or alternative tag to monitor the time progression of the phage escape from host.
IV assembly Modify in a way that increases p4 affinity for p1. This makes the channels more effective.
V binds ssDNA Modify such that p5 can sequester the + stranded DNA more effectively so that there is less competition with the formation of double stranded DNA.
VI phage tail protein (5 copies) Since p6 is the accessory protein to p3, it can be modified so that p3 can interact more effectively with the host.
VII phage head protein (5 copies) Since p7 is the companion protein of p9, make same modifications as that of p9.
VIII phage coat protein (2700 copies) Add myc or alternative tag to aid in targeting various types of hosts.
IX phage head protein (5 copies) Modify in such a way to make the phage secretion occur at a faster rate so that interaction time with the host is reduced. Also, modify so that p9 interaction with p5 is more effective. Since part of g9 overlaps with g8, be careful in making modifications.
X replication Modify such that the + strands of DNA are not soley dependent on the presence of p10. This modification works together with our modification of p2.
XI assembly Same modification as g1.