中文摘要：

目的原核表达及纯化重组果蝇jumeaux（jumu）蛋白,制备大鼠抗果蝇jumu多克隆抗体。方法用RT-PCR方法扩增jumu基因,并将其克隆到原核表达载体pRSETA中,转化大肠杆菌BL21（DE3）并用IPTG诱导His-jumu融合蛋白的表达。表达产物经SDS-PAGE及Western blot方法鉴定基因片段的正确性及蛋白表达的特异性后,用Ni-NTA亲和层析柱纯化带有6×His标签的融合蛋白,用纯化后的蛋白免疫SD大鼠制备多克隆抗体。以Western blot、免疫荧光染色法鉴定抗体的特异性,并用制备的抗体检测jumu在果蝇不同器官中的表达情况。结果成功构建了pRSETA-jumu原核表达质粒,jumu融合蛋白在大肠杆菌内高表达并纯化,免疫SD大鼠得到抗jumu多克隆抗体。用Western blot和免疫荧光染色检测发现制备的抗体有较强的特异性,并能检测内源性蛋白。通过免疫荧光染色发现jumu的表达存在组织特异性,且在唾液腺中的表达量最高。结论成功获得了抗jumu蛋白的特异性抗体,为进一步研究jumu蛋白的功能和作用机制奠定了基础。

英文摘要：

To purify the recombinant Drosophila jumeaux（jumu） protein and prepare rat polyclonal antibodies against jumu,we cloned a Drosophila jumu cDNA fragment and expressed it in BL21（DE3）.The expression products were purified by Ni-NTA affinity chromatograph,and then used to immunize SD rat to generate polyclonal antibodies.The specificity and the titer of the antibodies were detected by Western blotting;subcellular localization and tissue specific expression of jumu protein were analysed by immunostaining and immunohistochemistry.The results indicated that polyclonal antibodies against jumu could recognize the recombinant protein and native Drosophila jumu protein.Furthermore,analysis of immunostaining and immunohistochemistry showed that jumu protein was localized in the nucleus and expressed at high levels in salivary gland.In this study we successfully prepared recombinant 6×His-jumu protein and anti-jumu polyclonal antibodies with high titer and specificity,which may lay a basis for further study on the biological function of jumu protein.