Page 6 - V. Proteomics
P. 6
[V. Proteomics-6]
Proteome discovery of ASFV infected PAM cell
GaSeul Lee¹, DaeGwin Jeong², JeongHee Moon¹˙*
¹Disease Target Structure Research Center, KRIBB, Daejeon 34141, Korea, ²Infectious Disease Research Center,
KRIBB, Daejeon 34141, Korea
The differently expressed proteome by infection of African swine fever virus was investigated using porcine aveolar
macropahrge (PAM) cell. PAM cell was extracted from healthy swine lung and ASFV was infected in the BSL4 lab.
The noninfected PAM cell was also culutred as a control group. At the time point of 6h, 12h, 24h, 48h, 72h, both
the virus infected and noninfected cells were lysed with 5% SDS. Then, the lysed proteins can be brought out from
BSL4 lab safely.
The lysed proteins were digested with trypsin/lysC using S-trap. Then, peptides were labeled with TMT 11plex(126,
127C/N~131C/N), mid-pH RP fractionated and concatenated to 24 samples. The LC/MS/MS analysis conditions are
as follows : HCD mode, double play top 15 dda, 50cm C18 column, 7-60% ACN gradient 120min, 0.3ul/min flow
rate.
The peptide identification and reporter ion quantification was done with maxquant and perseus software. Swine
and ASFV database was used for identification. Heat map and hierachical clustering. The proteins which show similar
pattern with virus related proteins could be selected.
