User:Slokwong
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Name: Sophie Wong
Major: Course 20
Email: slokwong AT mit DOT edu
MODULE 1 : Genome Engineering
Notes on M13 Renovation
M13 genome
- 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)
- Independent genes:
- g3, g5, g6
Restriction enzymes used
- Pst1 digest
M13K07 :
Gene | Function | Idea |
---|---|---|
I | assembly | |
II | replication | modify such that it also nicks the other strand of the DNA to slow down the formation of double stranded DNA, thus reducing competition for p5. |
III | phage tail protein (5 copies) | add myc or alternative tag to monitor the time progression of the phage escape from host |
IV | assembly | |
V | binds ssDNA | modify such that it 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 it 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) | see modification for p9 since it is the companion protein. |
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 host is reduced. Also, modify it so that it interaction with p5 is more effective. Since part of g9 overlaps with g8, be careful in modification. |
X | replication | |
XI | assembly |