松材线虫纤维素酶的分离纯化及免疫学检测方法的研究  

Purification and Application of Bursaphelenchus Xylophilus Cellulase for Immunological Determination

作　　者：张奇[1] 马洪周[1] 杨文博[1] 白钢[1] 熊惠龙[2] 李海燕[2] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]国家林业局森林病虫害防治总站,辽宁沈阳110034

出　　处：《南开大学学报（自然科学版）》2006年第1期95-99,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家"十五"重点科技攻关子项目"林业重大病虫害预警及控制技术研究"

摘　　要：松材线虫分泌的纤维素酶在松树疫木的病理学研究上有着重要的地位．本研究通过松材线虫分泌的纤维素酶的鉴定以及初步纯化,得到了一种纤维素酶抗原(BXC60)．纯化后的纤维素酶抗原具有较强的水解羧甲基纤维素钠的能力,比活力为100．27 U／mg,与纯化前的粗酶液比活力相比提高了10．4倍,同时具有良好的抗原性．以该纤维素酶为指标,建立了一种选择性抗体酶联免疫分析法(SAEIA),该方法显示了较好的特异性,其检测灵敏度为0．1μg／well．利用该方法可以简便、快捷地检测出疫木中存在的纤维素酶,为松材线虫的检测及松材线虫病的研究提供了一种新的手段．The cellulase secreted from the Bursaphelenchus xylophis acted important roles in the process of pinewood infection. After preliminary identification and purification a specific cellulase antigen （BXC60） was gained from Bursaphelenchus xylophis. The BXC60 possess a stronger power in hydrolyzing CMC（specilfic activity： 100. 27 U ·mg ^-1）, and also have good antigenity, was purified 10.4-fold than crude enzyme extraction. Taking the BXC60 as target antigen, a selected antibody enzyme immunology assay （SAEIA） was established. The SAEIA method with satisfying specificity and the detection sensitivity is 0. 1 μg/well^-1. The method was able to identify the cellulase hidden in the infected pinewood simply, and showed a new potential for quarantine and studying of Bursaphelenchus xylophis.

关 键 词：松材线虫 纤维素酶 选择抗体酶联免疫分析法 

分 类 号：Q814.1[生物学—生物工程]