ELISA在苏云金杆菌杀虫蛋白毒素检测中的应用及新进展

doi:10.7668/hbnxb.2007.S2.051

华北农学报 ›› 2007, Vol. 22 ›› Issue (S2): 216-219. doi: 10.7668/hbnxb.2007.S2.051

ELISA在苏云金杆菌杀虫蛋白毒素检测中的应用及新进展

吴瑞华^1,2, 李国勋³, 冯书亮², 王容燕², 王金耀², 曹伟平², 杜立新², 陈丹²

1. 河北农业大学植物保护学院, 河北保定 071001;
2. 河北省农林科学院植物保护研究所, 河北保定 071000;
3. 青岛农业大学, 山东青岛 266109

收稿日期:2007-04-30 出版日期:2007-12-31
作者简介:吴瑞华(1981-),女,河北廊坊人,硕士,主要从事害虫生物防治研究
基金资助:
国家“863”计划项目(2006AA02Z189);河北省自然科学基金项目(C2005000642);河北省农科院青年基金项目(A06050101)

Application and the New Development of the Enzyme-linked Immunosorbent Assay to the Detections of Bacillus thuringiensis Insecticidal Crystal Protein

WU Rui-hua^1,2, LI Guo-xun³, FENG Shu-liang², WANG Rong-yan², WANG Jinyao², CAO Wei-ping², DU Li-xin², CHEN Dan²

1. College of Plant Protection,Agricultural University of Hebei,Baoding 071001,China;
2. Institute of Plant Protection,Hebei Academy of Agriculture and Forestry Sciences,Baoding 071000,China;
3. Agricultural University of Qingdao,Qingdao 266109,China

Received:2007-04-30 Published:2007-12-31

摘要/Abstract

摘要： 苏云金杆菌是目前应用较广泛的生物杀虫细菌之一,其产生的杀虫蛋白毒素是主要的杀虫成分,目前对苏云金杆菌的毒力和杀虫蛋白毒素的检测主要采用生物测定、SDS-PAGE等方法,但这些方法的灵敏度较差。本文综述了酶联免疫吸附分析法在苏云金杆菌蛋白毒素检测中的应用,以及在检测中采用的ELISA的类型和基本原理,并对其应用前景进行了展望。

关键词: 苏云金杆菌, 杀虫毒素蛋白, 酶联免疫吸附分析

Abstract: Bacillus thuringiensis(Bt) as one of the most widely used microbial pesticide,is a gram-positive bacterium that produces insecticidal proteins as crystalline protoxin inclusions during sporulation,and the Bacillus thuringiensis insecticidal protoxin is the main insecticidal ingredient,and the traditional detecting method of the insecticidal protoxin are bioassay and sodium dodecyl sulfate polyacrylamide gel electrophorasis(SDS-PAGE),but the methods are very complicated,at the same time not very sensitive.So the detecting method need to be improved.In this paper,applications of the Enzyme-linked Immunosorbent Assay(ELISA)to the detections of the Bacillus thuringiensis toxins,types and basic principles of the ELISA detected toxins from the Bacillus thuringiensis were all summarized,and the prospect of the ELISA was also reviewed.

Key words: Bacillus thuringiensis, Insecticidal crystal protein, Enzyme linked immunsorbsent assay

中图分类号:

S476.11

吴瑞华, 李国勋, 冯书亮, 王容燕, 王金耀, 曹伟平, 杜立新, 陈丹. ELISA在苏云金杆菌杀虫蛋白毒素检测中的应用及新进展[J]. 华北农学报, 2007, 22(S2): 216-219. doi: 10.7668/hbnxb.2007.S2.051.

WU Rui-hua, LI Guo-xun, FENG Shu-liang, WANG Rong-yan, WANG Jinyao, CAO Wei-ping, DU Li-xin, CHEN Dan. Application and the New Development of the Enzyme-linked Immunosorbent Assay to the Detections of Bacillus thuringiensis Insecticidal Crystal Protein[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(S2): 216-219. doi: 10.7668/hbnxb.2007.S2.051.

http://www.hbnxb.net/CN/Y2007/V22/IS2/216

参考文献

[1] 喻子牛.苏云金杆菌[M].北京:科学出版社,1990.
[2] 罗绍彬,阎建平,戴顺英,等.酶联免疫吸附测定(ELISA)检测苏云金杆菌伴胞晶体蛋白质研究[J].生物防治通报,1987,3(4):145-148.
[3] Schnepf N,Crickmore J,Van Rie Lereclus,et al.Bacillus thuringiensis and its pesticidal crystal protein[J].Microbiol And Molecular Biology,1998,62:775-806.
[4] Engvall E,Perlmann P.Enzyme-linked immunosorbent assay (ELISA)[C]//Quantitative Assay of Immunoglobulin G Immunochemistry,1971,8:871-874.
[5] Van Weeman B K,Schuurs A H W M.Immunoassay using Antigen-Enzyme conjugate[J].FEBS Letters,1971,15 (2):232.
[6] 焦奎,张书圣.酶联免疫分析技术及应用[M].北京:化学工业出版社,2004.
[7] Wie S I.Enzyme linked immunosorbent assay for detection and quantitation of the entomocidal parasporal crystalline protein of Bacillus thuringiensis subsp.kurataki and israelensis[J].Appl Environ Mlcrobiol,1982 43:891-894.
[8] Smith R A,Ulrich J T.Enzyme-linked immunosorbent assay for quantitative detection of Bacillus thuringiensis crystal protein[J].Appl Environ Microbiol,1983,45:586-590.
[9] 洪华珠,李星.酶联免疫吸附测定方法检测苏云金杆菌晶体蛋白质的研究[J].华中师范大学学报,1988,22(1):61-64.
[10] 沈鞠群,杨团员,喻子牛.酶联免疫吸附测定法测定四种苏云金杆菌菌株的晶体蛋白含量[J].生物防治通报,1990(增刊):47-50.
[11] Schnepf H E,Whiteley H R.Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli[J].Proc Natl Acad Sci USA,1981,78:2893-2897.
[12] 吴刚,崔海瑞,舒庆尧,等.Bt杀虫晶体蛋白基因及其转化基因育种研究进展[J].生物工程进展,2000,20(2):45-48.
[13] 李杨,任改新.苏云金杆菌Cry基因的遗传分析及基因工程[J].微生物学通报,1996,23(1):37-43.
[14] 张宏宇,邓望喜,喻子牛.苏云金杆菌的遗传多样性杀虫晶体蛋白基因型[J].遗传,2000,22(2):125-128.
[15] Hidetaka Hori,Yoshiyuki Takahashi,Makoto Takahashi,et al.Detection of the Bacillus thuringiensis serovar japonensis strain Buibui protoxin with enzyme-linked immunosorbent assay and its application to detection of the protoxin in soil[J].Appl Entomol Zool,2000,35(3):401-411.
[16] Tapp H,Stotzky G.Dot blot Enzyme-linked immunosorbent assay for monitoring the fate of insecticidal toxins from Bacillus thuringiensis in soil[J].Applied and Environmental Microbiology,1995:602-609.
[17] 沈法富,于元杰,尹承佾,等.利用Dot-ELISA检测Bt棉杀虫蛋白的研究[J].中国农业科学,1999,32(1):15-19.
[18] Kohler G,Milsten C.Continuous cultures of fused cells secreting antibody pre-defined specificity[J].Nature,1975,256:495.
[19] 王保民,何钟佩,田晓莉.苏云金芽孢杆菌杀虫晶体蛋白Cry1A单克隆抗体的制备及在转Bt基因棉毒蛋白检测上的应用[J].棉花学报,2000,12(1):34-39.
[20] 朱路青,曹越平.转Bt基因大豆植株种Bt毒蛋白的表达[J].上海交通大学学报(农业科学版),2005,23(3):234-238.
[21] Guesdon J L.The use of avidin-biotin interaction in immunoenzymatic techniques[J].J Histochem Cytochem,1979,27(2):3.
[22] Palm C J,Donegan K,Harris D,et al.Quantification in soil of Bacillus thuringiensis var.kurstaki delta-endotoxin from transgenic plants[J].Molec Ecol,1994,3:145-151.
[23] Tuan S J,Kao S S,Hsuan S L.Use of sodium dodecyl sulfate-ployacrylamide gel electrophoresis to quantify Cry1A protein of Bacillus thuringiensis subsp.kurstaki cell extract[J].J Plant Prot,1993,35:139-148(in Chinese).

选择文件类型/文献管理软件名称

选择包含的内容

ELISA在苏云金杆菌杀虫蛋白毒素检测中的应用及新进展

Application and the New Development of the Enzyme-linked Immunosorbent Assay to the Detections of Bacillus thuringiensis Insecticidal Crystal Protein

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 8

编辑推荐

Metrics

本文评价

[1]	燕树锋, 铁双贵, 岳润清, 韩小花, 齐建双, 卢彩霞. *转Cry1Ac-w基因抗虫玉米的获得及分子鉴定*[J]. 华北农学报, 2015, 30(5): 41-44.
[2]	王伟, 郭巍, 徐大庆, 孙伟明, 刘大群, 李瑞军. *苏云金杆菌cry8Ea2蛋白的分离纯化及生物活性测定*[J]. 华北农学报, 2012, 27(6): 48-52.
[3]	冯静静, 郭巍, 刘廷辉, 孙永祥, 孙伟明, 徐大庆. 苏云金杆菌cry7Ab8基因的克隆表达及其杀虫活性[J]. 华北农学报, 2011, 26(2): 35-38.
[4]	宋萍, 郭丽伟, 苏俊平, 王勤英. －株含cry新基因的苏云金杆菌菌株的研究[J]. 华北农学报, 2011, 26(1): 54-57.
[5]	王容燕, 冯书亮, 范秀华, 曹伟平, 胡明峻. 对金龟子幼虫有杀虫活性的苏云金杆菌HBF-1菌株发酵培养基优选[J]. 华北农学报, 2003, 18(S1): 96-98.
[6]	郭亚平, 马恩波, 任竹梅, 范仁俊, 薛锐. 苏云金杆菌制剂防治细胸金针虫的初步研究[J]. 华北农学报, 2001, 16(2): 108-112.
[7]	尹新明, 郭线茹, 贾新成, 吴云汉, 邱立友. 提高苏云金杆菌对棉铃虫毒力的研究[J]. 华北农学报, 1999, 14(2): 67-72.
[8]	冯书亮, 付韵琴, 范秀华, 王容燕, 邢建民, 胡明峻. 苏云金杆菌毒力指示昆虫玉米螟与棉铃虫、粘虫、黄地老虎的毒力相关性测定[J]. 华北农学报, 1995, 10(2): 91-94.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

ELISA在苏云金杆菌杀虫蛋白毒素检测中的应用及新进展

Application and the New Development of the Enzyme-linked Immunosorbent Assay to the Detections of Bacillus thuringiensis Insecticidal Crystal Protein

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 8

编辑推荐

Metrics

本文评价