Ideas 8
Ideas 8

1- Effect of salt, higher Ts on virion proteins (sharpen slightly)

2- Effect of salt, higher Ts on protease sensitivity of PBCV

3- Effect of incubating virions w/lysin on virion proteins

4- Effect of higher T incubation on lysin proteins

5- Protein composition of different lysin preps

6- Effect of heat on capsid protein in the absence of SDS

7- Can antibodies ppt PBCV from dilute solution (for SDS gel electrophoresis)

8- Develop conditions for proteolysis (minimize protease, inhibit w/PMSF)

9- Sensitivity of lysin to proteolysis

10- Improve lysin purif, LiCl in sucrose gradient, lower LiCl in pellet extraction, acid pH

10a- LiCl extraction series (sucrose pellet)

11- Gel filtration of better lysin preps

12- Wash PBCV w/detergents, high salt

12a- Use sarkosyl, pellet through sucrose pad

13- Deglycosylate capsid peptides

14- What proteins are at top of sucrose gradient?

15- Better gel on capsid peptides

16- Fresh lysin prep

17- Acid purification of PBCV, add increasing amounts of pH 4.5 buffer

18- Add LiCl systematically to PBCV cracking buffer