蚯蚓纤溶酶PI_(239)基因在杆状病毒中的表达

利用基因重组技术将PI2 39基因通过细菌 /杆状病毒 (Bac to Bac)表达系统构建重组杆状病毒表达载体 ,载体采用苜蓿银纹夜蛾核型多角体病毒 ,将重组病毒转染昆虫Sf9细胞 ,细胞出现明显病变。SDS PAGE电泳可以见到相对分子质量 3 .0万位置有蛋白表达 ,免疫印记结果表明该条带可以与PI2 39抗体反应。提示蚯蚓纤溶酶PI2 39重组蛋白已经获得表达 ,为进一步研究蚯蚓纤溶酶PI2 39基因的生物学活性奠定了基础

Expression of Lumbrukinase PI239 Gene in the Insect Cell using Baculovirus Expression Vector System

The objective of the study was to better understand the thrombolytic enzyme in earthworm (lumbrukinase). By using the baculovirus expression vector system, PI 239 protein was expressed in the insect cell. The PI 239 gene was inserted into the donor plasmid pFastbac. Through transposition, recombinant bacmid PI 239 DNA was formed and was then transfected into Sf9 insect cells to produce recombinant Baculovirus. Sf9 insect cell were infected with recombinant Baclovirous. The harvested culture supernatant and cells lysate occured after 72 h, the SDS PAGE and Western Blot analysis. The SDS PAGE analysis revealed that molecular weight of expressed protein was 30 000. Western blot showed that PI 239 protein could be recognized by specific antiPI 239 antibody. The PI 239 protein can be expressed on the insect cell Sf9.