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Cloning and Expression of BTB/POZ-Domain from Zebrafish gcl (Germ Cell-Less), and Appraisement of Its Polyclonal-Antibody

Cloning and Expression of BTB/POZ-Domain from Zebrafish gcl (Germ Cell-Less), and Appraisement of Its Polyclonal-Antibody
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摘要 In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl (germ cell-less) into the expression vector pET28c, and pET28c-gBTB/POZ was transformed into BL21(DE3) pLysS strain to express the fusion protein for the preparation of antibody. Polyclonal-antibody against the GCL-BTB/POZ domain was prepared by immunizing rabbit with the fusion protein, and the Western Blot and immuno-histochemical analysis were performed to detect the quantity of the polyclonal-antibody. The result indicates that the polyclonal-antibodies were of good quantity and specification. Further studies will be performed to demonstrate the function and expression pattern of the GCL protein during the development process of zebrafish with the polyclonal-antibody. In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl (germ cell-less) into the expression vector pET28c, and pET28c-gBTB/POZ was transformed into BL21(DE3) pLysS strain to express the fusion protein for the preparation of antibody. Polyclonal-antibody against the GCL-BTB/POZ domain was prepared by immunizing rabbit with the fusion protein, and the Western Blot and immuno-histochemical analysis were performed to detect the quantity of the polyclonal-antibody. The result indicates that the polyclonal-antibodies were of good quantity and specification. Further studies will be performed to demonstrate the function and expression pattern of the GCL protein during the development process of zebrafish with the polyclonal-antibody.
出处 《Wuhan University Journal of Natural Sciences》 CAS 2008年第1期123-128,共6页 武汉大学学报(自然科学英文版)
基金 Supported by the National Natural Science Foundation of China (30570968, 30370744)
关键词 gel (germ cell-less) polycolonal-antibody BTB/ POZ domain zebrafish (Danio rerio) gel (germ cell-less) polycolonal-antibody BTB/ POZ domain zebrafish (Danio rerio)
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