果蝇血液标记基因srp原核表达质粒构建及多克隆抗体制备

Prokaryotic Plasmid Construction and Polyclonal Antibody Preparation of Hematopoiesis Marker Gene Srp in Drosophila

果蝇基因srp是血液发育的早期标记基因,为了研究果蝇这一模式动物中血液发育的详尽过程,通过提取野生型成体果蝇的总RNA,反转录获得其cDNA文库,设计引物克隆出srp基因,将所得片段插入原核表达载体pET28a中,经酶切和序列鉴定,成功构建重组质粒,并通过IPTG诱导表达出His-srp融合蛋白,经Ni-IDA凝胶柱纯化后,免疫新西兰大白兔制备srp多克隆抗体,为血液发育研究奠定了基础.

Gene srp is a hematopoiesis marker in Drosophila. Reserve transcriptional PCR was performed using isolated total RNA from adult wild type. The part fragment of srp cDNA sequence was obtained by PCR amplification, cloned into prokaryotic expression pET-28a vector and identified with restriction enzyme and sequencing. The fusion protein was induced by IPTG, then purified by Ni-IDA gel column, and finally injected into New Zeal- and white rabbits to generate polyclonal antibody, which lays a foundation for blood developmental study to illuminate the elaborate process of hematopoiesis in Drosophila.