抗果蝇zip基因产物抗体的制备和胚胎zip蛋白分布的研究

PREPARATION OF ANTIBODIES AGAINST ZIP PRODUCT AND THE DISTRIBUTION OF ZIP PROTEIN IN DROSOPHILA EMBRYOS

zip基因为果蝇晚期神经发生所必需。分子生物学研究表明,zip基因产物是一个膜上整合糖蛋白,可能作为神经细胞的识别或粘联分子参与神经系统的发生。通过基因工程的方法,我们提取了lacZ-zip融合蛋白,继而免疫兔子制备了抗lacZ-zip融合蛋白抗体。该抗体在经过蛋白质印迹鉴定后,用于整幅果蝇胚胎的标记。结果显示zip基因产物主要在胚带缩短后表达,表明zip基因可能参与了晚期神经的发生。抗zip抗体除了识别中枢神经系统(CNS)中的个别神经元外,还标记了侧神经纤维,证实了以前的推测,即在CNS中表达的zip基因可能参与神经纤维束化的建立和维持。

Zip gene is required during the late neurogenesis of Drosophila. Molecular analysis of zip gene revealed that it encodes an integral membrane glycoprotein, functioning as a cellular recognition and/or adhesion molecule. It has been cloned into an expression plasmid pWR 590. By expression of this plasmid in E. coli, a fusion protein of zip with lacZ has been purified and used to prepare polyclonal antibodies from rabbits. After being identified by West blotting, antibodies were used to react with the whole mount embryos of Drosophila. The results of this antibody labelling experiment showed that zip proteins are fundamentally synthesized after germ band shortening, supporting our previous predication that zip gene is involved in the late neurogenesis and that antibodies against zip recognize lateral neural fascicles as well as neurons in CNS, proving that zip protein expressed in CNS may play a role in the establishment and maintenance of neural fasciculation.