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PLANT EXPRESSIONS OF CYTOKINES AND IIST APPILICATIONS FOR ANIMAL DISEASE CONTROL

细胞因子的植物表达和 IIST 在动物疾病控制中的应用

基本信息

批准号：
10556067
负责人：
SUGIMOTO Chihiro
金额：
$ 8.19万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10556067/
关键词：
INTERFERON TRANSGENIC PLANTS CYTOKINE PLANT VIRUS VECTOR

项目摘要

The aim of this study is to express bioactive animal proteins in plants and to apply the recombinant proteins for animal disease control. We used transgenic plant and plant viral vector stems for expression of animal genes.By Agrobacterium-mediated transformation, we introduced cDNAs of two subtypes of human interferon α (IFNα2b and IFNα8). Expression of the introduced genes was confirmed by northern blot analysis and enzyme immunoassay. Ten to 500 IU/g of IFN8 were detected in transgenic potatoes.In bovine leukemia virus infection, tumor necrosis factor α was revealed to be involved in pathogenesis, especially control of viral burden in infected animals. We also clones genes for a tick salivary grand protein, tick serine proteases, and a piroplasm surface antigen that are expected to be expressed in plant and used for method of disease controls.A plant expression system based on clover yellow vein virus has been developed by combining its cDNA and 35S promoter as pC1YVV. Expression of green fluorescence protein by this virus vector was confirmed. INF8 cDNA was introduced into this vector and its expression was tested. However, plant infected with this recombinant virus did not express any product as tested by enzyme immunoassay, provably because the viral vector had not retained the cDNA. Further investigation will be required for stable expression of animal genes in this system.

本研究的目的是在植物中表达具有生物活性的动物蛋白，并将重组蛋白应用于动物疾病的控制。利用转基因植物和植物病毒载体茎表达动物基因，通过农杆菌介导的转化，将人α干扰素（IFNα2b和IFNα8）的两个亚型的cDNA导入到植物中。通过北方印迹分析和酶免疫分析证实导入基因的表达。在转基因马铃薯中检测到10 ~ 500 IU/g的IFN-8。在牛白血病病毒感染中，肿瘤坏死因子α参与了致病过程，尤其是控制感染动物的病毒负荷。本研究还克隆了蜱类唾液大蛋白、蜱类丝氨酸蛋白酶和蜱类原生质体表面抗原的基因，以期在植物中表达并用于疾病控制。将其cDNA与35 S启动子结合构建了以三叶草黄脉病毒为载体的植物表达系统pC 1 YVV。证实了该病毒载体表达绿色荧光蛋白。将INF 8 cDNA导入该载体并检测其表达。然而，用该重组病毒感染的植物不表达任何产物，如通过酶免疫测定法所测试的，这可证明是因为病毒载体没有保留cDNA。动物基因在该系统中的稳定表达需要进一步的研究。