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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. |