脱落酸受体及其基因的分子免疫学研究(英文) |
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引用本文: | 周燮 金振华. 脱落酸受体及其基因的分子免疫学研究(英文)[J]. 安徽大学学报(自然科学版), 1998, 22(4): 79-85 |
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作者姓名: | 周燮 金振华 |
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作者单位: | 南京农业大学农学系,中国科学院发育生物研究所 |
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摘 要: | 发展了研究脱落酸(ABA)结合蛋白的两类免疫探针。其一,用ABA-C1-BSA-Sepharose4B亲和层析柱纯化出ABA结合蛋白,聚丙烯酰胺凝胶电泳显示该蛋白分子量为56KD的一条带,它具有特异结合ABA的能力(Kd=2.0×10-9mol/L)。当用蛋白水解酶K水解该蛋白,并用1%琼脂糖凝胶电泳时,发现其中存在约300个核苷酸的rRNA分子。用识别ABA结合蛋白的抗体筛选cDNA表达文库,从200,000个独立噬斑中获得120个编码玉米17sRNA的cDNA克隆和1个cDNA编码结合蛋白的cDNA克隆(24cDNA)。24cDNA有1075个碱基对,含编码254个氨基酸的开放阅读框架。其二,制备了识别抗ABA单克隆抗体的独特型抗体(anti-Id),它具有模拟并竞争ABA的能力。用上述两类免疫探针定位了植物细胞中的ABA结合蛋白。发展了研究脱落酸(ABA)结合蛋白的两类免疫探针。其一,用ABA-C1-BSA-Sepharose4B亲和层析柱纯化出ABA结合蛋白,聚丙烯酰胺凝胶电泳显示该蛋白分子量为56KD的一条带,它具有特异结合ABA的能力(Kd=2.0×10-9mol/L)。当用蛋白水解酶K水解该蛋?
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关 键 词: | 脱落酸 受体 基因 分子免疫学 结合蛋白 |
An Immunological Approach to ABA Receptor and Its Gene |
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Abstract: | Abstract Two types of immunological probes, anti-ABBP Abs, have been developed. The purified ABBP from ABA-C 1-BSA-sepharose 4B column was identified by PAGE and appeared in one band of about 56KD, as well as showed a specific binding ability and a high affinity for ABA (Kd2.0×10 -9 mol/L). Une〖HJ*3/7]xpectedly, the existence of rRNA with a length of around 300 nucleotides could be found, when the ABBP was digested with proteinase K and identified by electrophoresis on an agarose gel(1%). As a result, about 120 cDNA clones coding maize 17s RNA and only one cDNA clone coding ABBP (24cDNA) were obtained from 200,000 separated phage plaques by the anti-ABBP pAbs. 24cDNA had 1075bp and contained an open reading frame coding 254 amino acids. The anti-idiotypic Ab raised against an ABA MAb showed the ability of either mimicking ABA or competing with ABA. The localization of ABBPs in plant cell was investigated. |
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