Works Done

• Time Course Trypsin Digestion of P135K• Purification of P135K mutant with no AS• Fluoresence of P135K with Hep & AS• Fluoresence of P135K with Hep & no AS• Troubleshooting methods of lysing after overexpression

Future Works• Purification of P135K in AS, buffer exchange to no AS, run iTC• Re-transformation?• Inclusion body refolding?

0 3 6 9 12 15 18 21 24 27

Time Course Trypsin Digestion of P135K at 0.5 mg/mL Trypsin

17 kD Band

1. Urea2. 2 M NaCl3. 1.5 M NaCl4. 800 mM NaCl5. 500 mM NaCl6. 250 mM NaCl7. Flow Through8. Supernatant9. Pellet10. PM

1 2 3 4 5 6 7 8 9 10

SDS gel of Purification of P135K in no AS

1. PM2. 1.5M NaCl no AS3. 2M NaCl no AS4. 1500 Elute w/ AS5. Fr. Press Sup6. Son + Lysozyme

Sup7. Son + Lysozyme

Pellet8. Sonication Sup9. Sonication Pellet10. Pellet

1 2 3 4 5 6 7 8 9 10

SDS gel of Lysing Methods after Overexpression of P135K