中文摘要：

运用RT-PCR技术从盘基网柄菌（Dictyostelium discoideum）总mRNA中克隆到了尿囊酸酶基因（allC），该基因编码区开放读框长1100bp，编码的蛋白约42kD．由于是在野生型和突变型细胞中差异表达的片段，表明该基因在盘基网柄菌多细胞发育中起到重要作用，因此将allC克隆人融合表达载体pET-32a（＋），在大肠杆菌E．coliBL21（DE3）中进行诱导表达带有6个组氨酸标签的尿囊酸酶（ALC）融合蛋白，经镍柱亲和层析，获得了电泳纯蛋白．用纯化的融合蛋白免疫新西兰大白兔，制备多克隆抗体．ELISA测得制备的抗ALC蛋白的多克隆抗体的效价可达1：64000，WesternBlotting检测证明该抗体有较强的针对ALC蛋白的专一性．这些数据表明重组质粒表达的ALC融合蛋白具有良好的抗原性，制备ALC的多克隆抗体有良好特异性和效价，能够满足针对ALC免疫印迹和细胞内定位检测等实验要求，为深入研究ALC蛋白在盘基网柄菌多细胞发育的功能作用提供了有力工具．

英文摘要：

The coding region of allC is obtained from total mRNA of Dictyostelium discoideum by RT-PCR. The results show that the length of allC is 1 100 bp with sequence analysis and co- ded protein is 42 kD. Becase it is a differentially expression fragments, indicating that the gene play an important role in multicellular development and allC was cloned into the fusion expres- sion vector pET-32a（ ＋ ） with 6 His tagged and expressed in E. coli BL21 （DE3） host cells, and then purified by Ni2 ＋ affinity chromatography column and fusion protein purified was used to im- mune the New Zealand rabbits for preparing polyclonal antibody. The fusion protein was suc- cessfully expressed and polyclonal antibody was also successfully obtained. The potency of the antibody was as high as 1 ： 64 000. The specificity of antibody was proved by Western Blotting analysis of expression product of allC. These data suggest ALC fusion protein has the good anti- genicity. The antibody with high titer and specificity was obtained in the satisfaction of Western blot and localization experiment request , which was the foundation to farther study the charac- ters of the allantoicase protein and their function during Dictyostelium discoideum development.