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- W2376714622 abstract "Objective To construct eukaryotic expression v ector of vascular basement membrane-derived multifunctional peptide(VBMDMP) and to measure the anti-angio genesis activity of VBMDMP in MAE and anti-tumor activity of its fusion protein i n Human prostate adenocarcinoma cells (PC-3) transplanted into male SCID mice. Methods GST-VBMDMP containing Snab I and Not I enzyme cutt ing si tus was obtained by polymerase chain reaction(PCR),then purified,digested,recovered and cloned into pPIC9K eukaryotic expression vector. The recombinant pPIC9K-GST-VBMDMP w as transformed into E.coli DH5a and amplified, which was confirmed by restriction e n donuclease digestion and sequencing,then transformed into Pichia pastoris. T he control of pPIC9K was transformed into Pichia pastoristoo. Using a single c olon y, inoculated in MGY liquid medium and growed at 30 ℃ in a shaking incubator. A t each of the time points: 24 h? 48 h?72 h?96 h?120 h?144 h?168 h and 192 h , transf ered 1ml of the supernatants to 1.5ml EP tube and freezed in -70 ℃. At the mean t ime, kept the volume of culture and added methanol to a final concentration of 0 .5% every 24 h to maintain the induction. After 8 d, the supernatants were analyzed by SDS-PAGE for protein expression with Coomassie brilliant blue. The anti-tumor activity of GST-VBMDMP was measured which was purified by GST 4B Col u mn in Human prostate adenocarcinoma cells (PC-3) transplanted into male SCID mi ce and Endothelial Tube Assay was produced.Results The expre ssion vector of VBMDMP gene were constructed successfully.We obtained purified GST-VBMDMP fusion prot ein with GST 4B Column.The primary tumor inhibition rate were 96.6%, 82.1%, 61 .2% with GST-VBMDMP 10mg/kg, 6 mg/kg,2 mg/kg respectively. The tumor metastasi s inhibition rate of PC-3 were 96.8 %,87.9 %,75.3% with GST-VBMDMP 10 mg/kg , 6 mg/kg,2 mg/kg respectively. The data were significant difference in contras t to control group. Conclusion VBMDMP have significant anti- angiogenesis activity and have biological activity of anti-tumor growth and anti-metastasis in PC-3 transplanted into male SCID mice." @default.
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- W2376714622 date "2005-01-01" @default.
- W2376714622 modified "2023-09-26" @default.
- W2376714622 title "Eukaryotic Expression of Vascular Basement Membrane-derived Multifun-ctional Peptide and Its Anti-tumor and Anti-angiogenesis Activity Assay" @default.
- W2376714622 hasPublicationYear "2005" @default.
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