抗性家蚕血液中与BmNPV抗性相关铜化合物的分离方法研究

Study on the Separation Methods of Copper Compounds Related to BmNPV Resistance in Hemolymph of Bombyx mori

[目的]探究家蚕血淋巴中与BmNPV抗性相关铜化合物的分离方法。[方法]选取对BmNPV具有抗性的家蚕品种和非抗性品种为研究对象,对它们进行添食BmNPV病毒处理。提取家蚕的血淋巴,运用SEC-ICP-MS对2组样品中Cu化合物进行分离和检测。[结果]筛选出优化的体积排阻色谱分离条件:Yarra TM SEC-2000分离柱,30 mmol/L Tris-醋酸为流动相(pH 7.4),流速0.5 mL/min。建立了分离与检测家蚕血液中Cu化合物的SEC-ICP-MS联用技术。BmNPV抗性和非抗性家蚕血液中Cu化合物主要的差异是位于7.6 min的组分。[结论]制备了葡聚糖G100凝胶柱分离并收集可能与BmNPV抗性相关的家蚕血液分离液,为家蚕金属蛋白的分离提供更简便有效的方法。

[Objective]To study the separation methods of copper compounds related to BmNPV resistance in hemolymph of Bombyx mori.[Method]B.mori cultivars with and without BmNPV resistance were selected as research objects to feed BmNPV virus.Cu compounds was extracting and determinated from hemolymph of two groups B.mori by SEC-ICP-MS.[Result]The optimal seperation conditions for size exclusion chromatography were as follows：YarraTM SEC-2000,30 mmol/L Tris-acetic acid（pH 7.4） as mobile phase, flow rate of 0.5 mL/min.SEC-ICP-MS technology for separating and detecting copper compounds in hemolymph of B.mori was established.The main differences of copper compounds between resistant cultivars of B.mori and non-resistant cultivars of B.mori was the compounds with the retention time of 7.6 min.[Conclusion]The separation liquid of hemolymph of Bombyx mori might be related with BmNPV resistance seperated and collected by glucan G100 gel column was prepared.The research can provide simpler and more effective methods for the seperation of metal proteins in B.mori.