水稻纤维素合酶多克隆抗体的制备和鉴定
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国家转基因生物新品种培育科技重大专项(2009ZX08009-119B)


Preparation and identification of polyclonal antibodies of cellulose synthase in rice
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    摘要:

    纤维素合酶是参与纤维素β-1,4-葡聚糖链延伸的主要催化亚基,为进一步研究水稻中纤维素合酶的作用机理,采用DNA重组技术,将纤维素合酶基因CesA1、CesA2、CesA3克隆入表达载体pGEX-4T-3,构建重组质粒。在大肠杆菌JM109中,经异丙基硫代半乳糖苷(IPTG)诱导后在原核细胞中成功表达了3种纤维素合酶的融合蛋白,通过谷胱甘肽巯基转移酶(GST)纯化系统获得融合蛋白,以此作为抗原免疫家兔,制备出效价高、特异性强的多克隆抗体。

    Abstract:

    Cellulose synthase is a critical catalytic subunit of the cellulose synthase complex.To study functions of CesA genes in rice,their antibodies were prepared.First,recombinant plasmids were constructed with cloning the cellulose synthase genes into the expression vector pGEX-4T-3.The recombinant plasmids were then transformed into E.coli JM109 and the recombinant proteins were obtained.The fusion proteins were purified with GST affinity chromatography system and injected to rabbit for generating polyclonal antibodies.Finally,all antibodies were identified by Western blotting analysis with specific bands and high activities.

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张苗苗,陶章生,陈婷婷,夏涛,彭良才,丰胜求.水稻纤维素合酶多克隆抗体的制备和鉴定[J].华中农业大学学报,2011,30(4):393-397

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  • 收稿日期:2011-03-21
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