山西农业大学学报（自然科学版）2024，Vol.44Issue(2)：63-70,8.DOI:10.13842/j.cnki.issn1671-8151.202309034
蜂毒PLA2原核表达及溶血作用研究
Bee venom PLA2 Prokaryotic expression and hemolytic activity study
摘要
Abstract
[Objective]Bee venom is a natural animal medicine with broad pharmacological effects and complex composition.Phospholipase A2(PLA2)is an important active ingredient and major allergen of bee venom,which can synergize with melittin to induce hemolysis.This study aimed to express and purify BvPLA2 protein in prokaryotic system and compare the hemolytic effects of bee venom(BV),melittin,and BvPLA2 through hemolysis tests.[Method]The constructed pET28a-BvPLA2 fu-sion expression vector was transformed into E.coli BL21 strain,and the protein expression was induced by IPTG at low tem-perature.The expression product was identified and purified to obtain the recombinant BvPLA2 protein.Whole blood from mice was collected and mixed with physiological saline to prepare a 2%red blood cell suspension,and different concentrations of BvPLA2 protein solution,bee venom,and melittin were added.The hemolysis rate was determined by enzyme-linked im-munosorbent assay.[Results]The expression product induced by IPTG showed a clear band at approximately 15 kDa,consis-tent with the theoretical calculation value.Specific bands were expressed in both the supernatant and sediment after ultrasonica-tion,and the expression levels in the supernatant were basically consistent with those in the sediment.BvPLA2 protein solu-tions at different concentrations exhibited hemolytic effects,with hemolytic rates exceeding 5%,and the hemolysis rate was higher at low concentration of BvPLA2.By comparison,the hemolysis rate of BvPLA2 was significantly lower than that of bee venom and melittin(P<0.05).[Conclusion]Soluble target protein BvPLA2 was successfully induced and purified through the prokaryotic expression system,and BvPLA2 exhibited certain hemolytic effects,but the hemolytic effect was significantly low-er than that of bee venom and melittin.关键词
蜂毒/磷脂酶A2/原核表达/溶血性Key words
Bee venom/Phospholipase A2/Prokaryotic expression/Hemolysis分类
生物科学引用本文复制引用
刘苗苗,王兆宇,陈龙龙,吕建华,赵慧婷..蜂毒PLA2原核表达及溶血作用研究[J].山西农业大学学报（自然科学版）,2024,44(2):63-70,8.基金项目
山西省自然科学青年基金项目(201901D211356) （201901D211356）
