使用5种分离培养基分离南方红豆杉〔Taxus wallichiana var.mairei(Lemée et H.Lév.)L.K.Fu et N.Li〕根、枝和叶中的内生真菌,并对分离的菌株进行分子鉴定;采用HPLC技术分析不同内生真菌处理组南方红豆杉叶中紫杉醇及中间产...使用5种分离培养基分离南方红豆杉〔Taxus wallichiana var.mairei(Lemée et H.Lév.)L.K.Fu et N.Li〕根、枝和叶中的内生真菌,并对分离的菌株进行分子鉴定;采用HPLC技术分析不同内生真菌处理组南方红豆杉叶中紫杉醇及中间产物巴卡亭Ⅲ和10-去乙酰基巴卡亭Ⅲ(10-DAB)的含量差异。结果表明:在南方红豆杉的根、枝和叶中共分离到69株内生真菌,隶属于26属36种;Muyocopron Speg.属有15株内生真菌,占分离的内生真菌菌株总数的21.7%,为南方红豆杉内生真菌的优势属。在19个内生真菌处理组南方红豆杉的叶中可同时检测到紫杉醇、巴卡亭Ⅲ和10-DAB,且这些处理组的巴卡亭Ⅲ和10-DAB含量均低于对照组(未喷洒内生真菌诱导子溶液),而紫杉醇含量却存在明显差异。其中,10个内生真菌处理组的紫杉醇含量高于对照组,以Pseudodidymocyrtis lobariellae Flakus,Rodr.Flakus et Etayo处理组的紫杉醇含量最高(0.228 mg·g^(-1)),Gymnopilus luteofolius(Peck)Singer和Coniochaeta hoffmannii(J.F.H.Beyma)Z.U.Khan,Genéet Guarro处理组的紫杉醇含量也较高(分别为0.180和0.139 mg·g^(-1))。重复验证结果表明:与对照组相比,P.lobariellae处理组南方红豆杉叶中紫杉醇含量显著升高,但巴卡亭Ⅲ和10-DAB含量明显下降。研究结果显示:南方红豆杉内生真菌多样性较高,部分内生真菌能够促进南方红豆杉叶中紫杉醇的合成,其中,P.lobariellae、G.luteofolius和C.hoffmannii可作为调控南方红豆杉紫杉醇积累的候选菌。展开更多
Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals, The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer m...Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals, The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene （ompW） from E parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 （DE3） cells. The gene coded for a protein that was 42.78 kDa. We purified the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for future application of the OmpW protein as a vaccine candidate against infection by E parahaemolyticus. In addition, the purified OmpW protein can be used for further functional and structural studies.展开更多
基金Supported by the Key Laboratory Foundation of the Educational Department of Liaoning Province (No. 2009S024)the Dalian Municipal Government of China (No. 2007B11NC069)the Grant of Dalian Ocean University (No. SY2007005)
文摘Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals, The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene （ompW） from E parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 （DE3） cells. The gene coded for a protein that was 42.78 kDa. We purified the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for future application of the OmpW protein as a vaccine candidate against infection by E parahaemolyticus. In addition, the purified OmpW protein can be used for further functional and structural studies.
