PREPARATION AND APPLICATION OF MONOCLONAL ANTIBODY TO SUGARCANE (SACCHARUM L. SPP. HYBRIDS) SOP5CS PROTEIN
文献类型： 外文期刊
作者： Li, J. 1 ; Wang, L. R. 1 ; Huang, X. 2 ; Xing, Y. X. 1 ; Yang, L. T. 1 ; Li, Y. R. 2 ;
作者机构： 1.Guangxi Univ, Coll Agr, Nanning 530005, Peoples R China
2.Guangxi Acad Agr Sci, Chinese Acad Agr Sci, Minist Agr & Rural Area, Key Lab Sugarcane Biotechnol & Genet Improvement, Nanning 530007, Peoples R China
3.Guangxi Acad Agr Sci, Chinese Acad Agr Sci, Guangxi Key Lab Sugarcane Genet Improvement, Nanning 530007, Peoples R China
4.Guangxi Acad Agr Sci, Chinese Acad Agr Sci, Sugarcane Res Ctr, Nanning 530007, Peoples R China
关键词： sugarcane; P5CS gene; proline; monoclonal antibody; physiological index
期刊名称：APPLIED ECOLOGY AND ENVIRONMENTAL RESEARCH （ 影响因子：0.711； 五年影响因子：0.796 ）
ISSN： 1589-1623
年卷期： 2019 年 17 卷 5 期
页码：
收录情况： SCI
摘要： Delta(1)-pyrroline-5-carboxylate synthase (P5CS) is a key enzyme in proline biosynthesis. In this study, the nucleotide sequence of sugarcane P5CS (SoP5CS) gene was obtained from sugarcane. Then the ORF of SoP5CS linked with pET-30a vector and the SoP5CS protein was expressed successfully in E. coli by the induction of 1.0 mM IPTG at 28 degrees C for 6 h. The prokaryotic expression protein was expressed as inclusion bodies (IBs). It was purified by Ni-NTA Resin firstly and then refolded, dialyzed and further purified by gel filtration. The monoclonal antibody of SoP5CS was prepared using mice. The expression of SoP5CS gene and the corresponding protein and physiological parameters associated with drought resistance were detected in sugarcane leaves under different drought stresses, they worked together to regulate the drought resistance process of sugarcane. This is the first report on the preparation of P5CS monoclonal antibody and the expression of the antibody in sugarcane, which would provide new methods to study the function of SoP5CS response to various abiotic stresses and other aspects.
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