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Updated: Aug 18, 2026
Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells
Published on: May 9, 2020
[Cloning, expression, purification and characterization of human endostatin gene]
Fang Yang1, Yuan-Li He, Xiao-Yu Jiang
1Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Objective:
To procure biologically active human endostatin.
Methods:
Human endostatin gene was acquired by means of reverse transcriptase (RT)-PCR and cloned into PGEM-T vector with subsequent sequence identification. The gene fragment was inserted into the prokaryotic expression vector pBV220 and transformed into E.coli DH5alpha strain. Endostatin expression in the E.coli was identified and the inclusion body isolated, purified and its activity analyzed.
Results:
The obtained gene fragment 552 bp in length was identified as the functional section of human endostatin gene by sequence analysis, and SDS-PAGE analysis showed that the expressed product was the target protein with biological activity.
Conclusion:
Human endostatin gene was expressed in E.coli and the protein obtained can inhibit the proliferation of ECV 304 cells.
