利用家蚕病毒颗粒表达和纯化抗肿瘤药物天冬酰胺酶

Expression and purification of antitumor drug L-asparaginase with Bombyx mori virus particles

目的:通过家蚕病毒颗粒简化抗肿瘤药物L-天冬酰胺酶的生产纯化过程,为抗肿瘤的药物生产提供依据。方法:利用基因重组技术构建pFastdual-polh-da26-asp融合蛋白真核表达载体,利用杆状病毒表达系统在家蚕细胞中表达融合蛋白,奈氏试剂法检测L-天冬酰胺酶活性。结果:成功构建了重组表达载体,融合蛋白在家蚕细胞中顺利表达,并检测到L-天冬酰胺酶在家蚕细胞中表达,活性为2 113.52 U/mg。结论:重组病毒颗粒成功表达,且L-天冬酰胺酶具有一定的活性;可以利用家蚕颗粒病毒简化纯化抗肿瘤药物L-天冬酰胺酶。

Objective： To simplify the production and purification process of antitumor drug L-asparaginase by Bombyx mori virus,and to provide the basis for drug production of tumor therapy.Methods： The pFastdual-polh-da26-asp fusion protien eukaryotic expression vector was constructed by gene recombination technique.The fusion protein was expressed by the baculovirus expression system in Bombyx mori cells.The activity of L-asparagine was detected by using nessler reagent.Results： The recombinant expression vector was constructed successfully;and the fusion protein was expressed in Bombyx mori cells.The activity of L-asparagine was 2 113.52 U/mg.Conclusion： The recombinant virus particles were expressed efficiently,and the product L-asparagine has certain activity.