Figure 2: Expression of tumstatin in E. coli and mammalian
cells
(A) Vector map of pET22b harboring tumstatin: Tumstatin was cloned between NdeI and XhoI sites of bacterial expression vector pET22b. (B) E. coli expressed solubile tumstatin protein: Lane 1, shows molecular weight marker; lane 2 shows about 10μl of column flow through; lane 3 and 4 show total soluble and insoluble tumstatin protein (total soluble and insoluble tumstatin protein suspended in equal volume of PBS and 2 μl was loaded in each lane). (C) Vector map of plasmid pcBFT harboring tumstatin: Tumstatin was cloned in frame between BamHI and XhoI sites of mammalian expression vector, pcBFT. (D) HEK- 293 cell expressed soluble tumstatin: Lane 1 is a molecular weight marker; lane 2 shows cytosolic extracts from HEK-293 cells; lanes 3 and 4 show soluble and insoluble tumstatin fractions similar to B. B and D are coomassie stained 10% SDS-PAGE gel.
(A) Vector map of pET22b harboring tumstatin: Tumstatin was cloned between NdeI and XhoI sites of bacterial expression vector pET22b. (B) E. coli expressed solubile tumstatin protein: Lane 1, shows molecular weight marker; lane 2 shows about 10μl of column flow through; lane 3 and 4 show total soluble and insoluble tumstatin protein (total soluble and insoluble tumstatin protein suspended in equal volume of PBS and 2 μl was loaded in each lane). (C) Vector map of plasmid pcBFT harboring tumstatin: Tumstatin was cloned in frame between BamHI and XhoI sites of mammalian expression vector, pcBFT. (D) HEK- 293 cell expressed soluble tumstatin: Lane 1 is a molecular weight marker; lane 2 shows cytosolic extracts from HEK-293 cells; lanes 3 and 4 show soluble and insoluble tumstatin fractions similar to B. B and D are coomassie stained 10% SDS-PAGE gel.