家蝇抗菌物质的电泳分离及其条件优化

目的 研究以针刺诱导家蝇产生抗菌物质,对家蝇针刺诱导后抗菌物质的电泳分离条件以及热处理方法进行初步的探讨.方法 将家蝇3龄幼虫针刺体壁诱导48 h后提取血淋巴,100℃水浴血淋巴0、0.5、1、3、5 min,分别取上清液5、10、20、30μl进行酸性聚丙烯酰胺凝胶电泳,以分离家蝇的抗菌物质.结果 热处理和未经热处理的血淋巴都以20μl的加样量分离效果较好,热处理时间以100℃水浴加热1 min血淋巴的电泳效果较好.比较针刺诱导和未经诱导家蝇的血淋巴,结果显示诱导后血淋巴中的蛋白质无论是种类还是含量上都有所增加,经过100℃水浴加热1 min后,血淋巴条带的数量和丰度均有所增加.结论 100℃水浴加热1 min,加样量为20μl为家蝇血淋巴最佳处理效果.针刺诱导可以使家蝇血淋巴产生新的蛋白质,这些蛋白质可能是诱导产生的新的抗菌物质.

Abstract：

Objective To study the antibacterial substances produced by Musca domestica and heattreatment and electrophoresis isolation of the substances. Methods After the 3rd instar larval of Musca domestica being induced for 48 hours by pricking, the hemolymph was collected and bathed at 100 ℃ for 0,0.5,1,3,5 min. Antibacterial substances of Musca domestica was isolated by electrophoresis with acide polyacrylamide gel using 5,10,20,30 μl doses. Results 20 μl was the optimum dose both for bathed and non-bathed hemolymph. Water bath at 100 ℃ for 1 min was the optimum bath time for hemolymph. Comparing induced hemolymph with non-induced hemolymph, the protein of the induced hemolymph was increased in the number and content. After 100 ℃ bathed for 1 min,it showed that the bonds of hemolymph increased both in quantities and contents. Conclusion Optimum treatment is 100 ℃ bath for 1 min and 20 μl of adding dose. Pricking inducing can produce new proteins in hemolymph, which are the new induced antibacterial substances.

Isolation of Antibacterial Sunstances from Musca domestica and Optimum of Electrophoresis

Objective To study the antibacterial substances produced by Musca domestica and heattreatment and electrophoresis isolation of the substances. Methods After the 3rd instar larval of Musca domestica being induced for 48 hours by pricking, the hemolymph was collected and bathed at 100 ℃ for 0,0.5,1,3,5 min. Antibacterial substances of Musca domestica was isolated by electrophoresis with acide polyacrylamide gel using 5,10,20,30 μl doses. Results 20 μl was the optimum dose both for bathed and non-bathed hemolymph. Water bath at 100 ℃ for 1 min was the optimum bath time for hemolymph. Comparing induced hemolymph with non-induced hemolymph, the protein of the induced hemolymph was increased in the number and content. After 100 ℃ bathed for 1 min,it showed that the bonds of hemolymph increased both in quantities and contents. Conclusion Optimum treatment is 100 ℃ bath for 1 min and 20 μl of adding dose. Pricking inducing can produce new proteins in hemolymph, which are the new induced antibacterial substances.