Harpin Expression and Induction of Bioactivity to Anti-TMV Infection

XU Jin-ping; MENG Xiao-lin; WANG Jian; LU Wei; ZHANG Jun-jie

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December 2003

XU Jin-ping, MENG Xiao-lin, WANG Jian, LU Wei and ZHANG Jun-jie. Harpin Expression and Induction of Bioactivity to Anti-TMV Infection[J]. Virologica Sinica, 2003, 18(6): 607-610.

Citation: XU Jin-ping, MENG Xiao-lin, WANG Jian, LU Wei, ZHANG Jun-jie. Harpin Expression and Induction of Bioactivity to Anti-TMV Infection .VIROLOGICA SINICA, 2003, 18(6) : 607-610.

Harpin的表达及其诱导抗烟草花叶病毒感染的活性

1.
武汉大学病毒研究所，湖北武汉，430072

摘要

摘要：采用人工分段合成梨火疫病细菌(Erwinia amylovora)Harpin基因A、B、C、D片段，然后连接构建全长 harpin基因，克隆到pET-23(+)表达载体中，在E．coliBL21中表达Harpin蛋白。通过His BindResin Ni” 层析柱纯化，获得Harpin蛋白。经SDS．PAGE和Westernblot分析，该蛋白分子量为45 kDa。采用叶片穿刺法，5pL3 edrnL Harpin能诱发烟草叶片的过敏反应。采用叶脉注射法，5pL 300g／mL Harpin能诱发辣椒叶片的过敏反应。将烟草和辣椒用3 g／mE Harpin喷雾处理5d后，接种烟草花叶病毒(Tobacco mosaic virus，TMV)，观察植株病症，并采用ELISA检测TMV含量。结果表明经Harpin处理的烟草和辣椒对TMV侵染有一定的抑制作用。
- Harpin蛋白：合成
- , 表达
- , 纯化
- , 烟草花叶病毒

Harpin Expression and Induction of Bioactivity to Anti-TMV Infection

1.
Instiute of Virology,Wuhan University,Wuhan 430072，China

Abstract

Abstract：Harpin gene A，B，C and D fragments of Erwinia amylovora(Ea)were synthesized．The full length Harpin gene was constructed by ligation，and then cloned into pET-23(+)．Th e fusion Harpin protein Was expressed in E．coli BL2 1，purified by His Bind Resin Ni”chromatography and subjected to SDS—PAGE an d Western blot．The Mr of the Harpin was 45 kDa．5pL 301ag／mE Harpin induced hypersensitive response on tobacco or capsicum leaves by using punctures or injection along nervation in laminae respectively．Tobacco an d capsicum were treated with 30pg／mE Harpin protein by spraying over the plant，and then Tobacco mosaic virus(TMV)was inoculated in the plants by mechanical inoc ulation after 5 days．Th e TM V concentration in leaves was determined by ELISA．TM V infection in tobacco an d capsicum treated with Harpin was repressed to a certain extent．
- Harpin protein
- , Syn thesize
- , Expression
- , Purification

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

Harpin Expression and Induction of Bioactivity to Anti-TMV Infection

1. Instiute of Virology,Wuhan University,Wuhan 430072，China

Abstract: Abstract：Harpin gene A，B，C and D fragments of Erwinia amylovora(Ea)were synthesized．The full length Harpin gene was constructed by ligation，and then cloned into pET-23(+)．Th e fusion Harpin protein Was expressed in E．coli BL2 1，purified by His Bind Resin Ni”chromatography and subjected to SDS—PAGE an d Western blot．The Mr of the Harpin was 45 kDa．5pL 301ag／mE Harpin induced hypersensitive response on tobacco or capsicum leaves by using punctures or injection along nervation in laminae respectively．Tobacco an d capsicum were treated with 30pg／mE Harpin protein by spraying over the plant，and then Tobacco mosaic virus(TMV)was inoculated in the plants by mechanical inoc ulation after 5 days．Th e TM V concentration in leaves was determined by ELISA．TM V infection in tobacco an d capsicum treated with Harpin was repressed to a certain extent．