Figure 10
From: Implications of high level pseudogene transcription in Mycobacterium leprae
Cloning and protein expression of the pyrR pseudogene in E. coli. Crude cell lysates of recombinant BL21-Star E. coli cells containing the M. leprae pyrR pseudogene (E. coli::pET200/D/TOPO/MLpyrR) were separated on 4–20% SDS-PAGE gradient gels, and the histidine epitope was detected by Western blotting using the Anti-Xpress™ antibody: Panel A, shows the M. leprae pyrR pseudogene sequence for PCR amplification, yellow bases depict primers for PCR, the numbered sequences are the coding region; Panel B, is an SDS-PAGE gel stained with commassie brilliant blue, containing proteins from the recombinant E. coli strains: Lane 1, Kaleidoscope Prestained Standards Invitrogen #161-0324; Lane 2, E. coli::pET200/D/lacZ; Lane 3, E. coli::pET200/D/lacZ, IPTG-induced 18 hr; Lane 4, E. coli::pET200/D-TOPO/MLpyrR; Lane 5, E. coli::pET200/D-TOPO/MLpyrR, IPTG-induced 18 hr. Panel C represents a Western blot for the histidine epitope on the recombinant E. coli::pET200/D-TOPO/MLpyrR using crude cell lysates from IPTG-induced 18 hr samples: Lane 1, E. coli::pET200/D/lacZ; Lane2, E. coli::pET200/D/TOPO/MLpyrR.