华支睾吸虫不同发育期同工酶及蛋白质的研究

目的通过对华支睾吸虫不同发育时期的同工酶及蛋白质分析,了解其变化规律。方法以浓度梯度-聚丙烯酰胺凝胶电泳(CG-PAGSE)对10、20、30、609、0 d五个虫龄期华支睾吸虫乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、脂酶(EST)、过氧化物酶(PO)、超氧化物歧化酶(SOD)进行同工酶分离,应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离蛋白质(PT),用凝胶成像软件对酶(蛋白)带的迁移率、相对浓度分布、光密度等结果进行分析。结果60 d、90 d虫龄期LDH-5相对浓度增加;MDH以90 d组酶带最多,为8条;EST以20 d3、0 d组酶带最多,分别为8条和7条;PO在60 d9、0 d组分别多了1条带;SOD酶带随虫龄增大呈减少趋势;PT分子质量在6.0~169.8 ku之间,随虫龄增大带数增加。结论华支睾吸虫不同发育时期的同工酶及蛋白质变化有一定规律,实验结果为该虫的发育学、生理、生化及杀虫药物研究等提供了基础资料。

Studies on the patterns of isoenzymes and proteins in different development stages of Clonorchis sinensis

Objective To reveal the transformation rule of the patterns of isoenzymes and proteins at different developmental stages of Clonorchis sinensis.Methods Concentration gradient polyacrylamide gel slab electrophoresis(CG-PAGSE) was carried out to isolate the isoenzyme from Clonorchis sinensis at ages 10 d,20 d,30 d,60 d,90 d,which include lactate dehydrogenase(LDH),malate dehydrogenase(MDH),esterase(EST),peroxidase(PO) and superoxide dismutase(SOD);and SDS-polyacrylamide gel electrophoresis(SDS-PAGE) was carried out to isolate the the general protein(PT).A gel image analysis software was applyed to analyse the relative mobility,relative protein concentration profile and optical density.Results The relative protein concentration of LDH-5 at ages 60 d and 90 d were more than other ages.The bands number of MDH at 90 d got the maximum 8.EST bands number were more than others at the stages of 20 d and 30 d,which were 8 and 7.There were one more band of PO at stages 60 d and 90 d.The bands number of SOD decreased when the worm grew.The protein molecular weight range from 6.0 ku to 169.8 ku.The PT bands were more and more as the worm grew.Conclusion Some variations were detected in the patterns of isoenzymes and proteins during C.sinensis' development.The results could provide basic datum for research of C.sinensis' development biology,physiology,biochemistry and pesticide.