Expression and Identification of Mutated Osteoprotegerin in Culture Cells and Larvae of Silkworm
Expression and Identification of Mutated Osteoprotegerin in Culture Cells and Larvae of Silkworm作者机构：Institute of Biochemistry College of Life Sciences Zhejiang University Hangzhou 310029 China College of Life Sciences Zhejiang University of Science & Technology Hangzhou 310018 China
出 版 物：《Acta Biochimica et Biophysica Sinica》 (生物化学与生物物理学报(英文版))
年 卷 期：2004年第36卷第5期
页 面：331-335页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种]
主 题：OPG-372 silkworm baculovirus expression system hypocalcemic effect
摘 要：The mutated osteoprotegerin (OPG-372) gene was inserted into the baculovirus transfer vector pBacPAK8, and the recombinant plasmid was co-transfected with linearized Bm-BacPAK6 virus DNA into BmN cells, then homologous recombination occurred inside the cells. The recombinant virus BmNPV-OPG- 372 was screened and identified by Southern blotting. The recombinant human OPG-372 was expressed in cultured cells and the larvae of silkworm by inoculation of recombinant virus. The expression products were run on the SDS-PAGE and their immunoreactivities were determined by Western blotting. It was found that a 42 kD recombinant protein was expressed in BmN cells and a 46 kD one in larvae respectively. The bioactivi- ties of the recombinant proteins were determined by hypocalcemic effect assay in the mice. The results showed that the recombinant proteins had a significant hypocalcemic effect on mice sera.
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