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Expression and Activity Determination of Human Angiostatin (k1-3) in Culture Cells and Larvae of Silkworm

Expression and Activity Determination of Human Angiostatin (k1-3) in Culture Cells and Larvae of Silkworm

Ying-Fei Wang, Yong-Feng Jin, Yao-Zhou Zhang

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2002 Jul;34(4):482-7.

PMID: 12098772

Abstract:

Angiostatin (k1-3) gene was inserted into Bombyx mori baculovirus transfer vector pBacPAK8 and cotransfected with lineared DNA of Bm-BacPAK6 virus into BmN cells. The homologous recombination occurred inside the cells, and the recombinant virus BacPAK-angiostatin was expressed, as identified by DNA dot blotting. The BmN cells and fifth instars were infected by the recombinant virus BacPAK-angiostatin; expression product was run in the SDS-PAGE, and its immunoreactivity was determined by using ELISA and Western blotting. The bio-activity of the protein product was determined by using human umbilical vein endothelial cells ( ECV304 ) proliferation test in vitro and by using CAM vascular inhibition test in vivo. The expression activity achieved the highest point at the 72nd hour in BmN cells (22 u / 2x10(6) cells) and at 144th hour in larvae (159 u/ml). At the concentration of 2.5 u/ml, angiostatin induced apoptosis of endothelial cells in 24 h and also inhibited angiogenesis in CAM.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413057	Angiostatin K1-3 human			Price

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