Analysis of the protein in hemolymph from Anopheles stephensi during melanotic encapsulation of Plasmodium yoelii oocysts by two-dimensional electrophoresis / 第三军医大学学报

ABSTRACT

Objective To analyze the protein in hemolymph from adult female Anopheles stephensi (An. Stepheni) infected by Plasmodium yoelii after feeding with sucrose solution containing nitroquine or simple sucrose solution with two dimensional gel electrophoresis technique. Methods Hemolymphs from nitroquine fed, infected blood fed, and sucrose solution fed adult female An. stephensi were collected using the expulsion method on the third day after the feeding. Hemolymph protein concentration was examined with Bradford method. Then the hemolymph protein was analyzed by two dimensional electrophoresis. The protein spots were visualized by Coomassie brilliant blue staining. The spots were scanned and automatically analyzed by the ImageMaster VDS CL (Amersham Pharmacia) and ImageMaster 2D Elite software (Amersham Pharmacia). Results The protein concentration in the nitroquine fed group was always lower than that in the infected blood fed and sucrose solution fed groups. Two dimensional gel electrophoresis revealed 101 protein spots in nitroquine fed and 115 protein spots in the control with 51 matched, but unmatched 50 and 64 protein spots were detected in the treatment group and the control group, respectively. Different protein spots were mainly located at the molecular weight of (40-60)?10 3 and at the isoelectric points of basic end. Conclusion Two dimensional gel electrophoresis may directly reflect the difference of the protein. Both the difference of protein concentration and the protein spots may be involved in nitroquine induced melanotic encapsulation of Plasmodium yoelii oocysts.