摘要
应用淋巴细胞杂交瘤技术,融合经水稻细菌性条斑病病原菌(Xanthomonas campestris pv.oryzicola)免疫的BALB/C小鼠脾细胞和Sp2/O骨髓瘤细胞,经细条病菌典型菌株R17,R25和GX-1单独及三者混合包被进行ELISA检测筛选,建立了能稳定分泌特异性单克隆抗体的杂交瘤细胞株18G 9,19D8,21F2,21G3,25E6和26F9细胞株。体外连续传代培养三个月,液氮冻存四个月后复苏培养,分泌抗体稳定。六株杂交瘤细胞的腹水单克隆抗体ELISA效价在1:2000左右。各株细胞的培养上清液ELISA效价为1:8左右。18G9,19D8和25E6细胞株所分泌的抗体为小鼠IgM类,21F2和21G3分泌的抗体为小鼠IgG_3亚类,26F9分泌的抗体为小鼠IgG_(2b)亚类。根据与植物病原细菌4个属6个种的20个不同菌株的特异性反应,将六株单克隆抗体分为A,B,C三组,20个菌株分为3个血清型和1个菌株群。
Six hybridoma cell lines (18 G 9, 19 D 8, 21 F 2, 21 G 3, 25 E 6 and 26 F 9 ) secreting the monoclonal antibodies (McAbs ) to Xanthomonas campsstris pv. oryzicola were obtained. The hybridomas still secreted stably the pathogenic bacterium specific McAbs after a serial culture in vitro and storage in liquid nitrogen. It was proved that the antibodies of the lines 18 G 9, 19 D 8 and 25 E 6 belong to murine IgM, 2l F 2 and 21 G 3 to IgG3, and 26 F 9 to IgG 2b . At least three epitopes and three serovars were supposed after thirteen strains of Xanthomonas were tested with the McAbs. Serovar Ⅰ, including strains R17, R 25 and GX-1, reacted with all McAbs of A, B, C group. Serovar Ⅱ, the strains R21, R 22, R 27, SI, S2, S3, S17, S24 and H4, reacted with the McAbs of B, C group; Serovar Ⅲ of the strain S20 reacted only with the McAbs of C group. The significance and problemsin the application of the McAbs were discussed.
关键词
水稻
条斑病
病原菌
单克隆抗体
bacterial leaf streak of rice
monoclonal antibody
serovar
