Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ciruna B, Weidinger G, Knaut H, et al. Production of Maternal-Zygotic Mutant Zebrafish by Germ-Line Replacement [J]. Proc Natl Acad Sci USA, 2002, 99(23): 14919–14924.
Jongens T A, Hay B, Jan L Y, et al. The Germ Cell-Less GeneProduct: a Posteriorly Localized Component Necessary for Germ Cell Development in Drosophila [J]. Cell, 1992, 70 (4): 569–584.
Kimura T, Yomogida K, Lwai N, et al. Molecular Cloning and Genomic Organization of Mouse Homologue of Drosophila Germ Cell-Less and Its Expression in Germ Lineage Cells [J]. Biochem Biophys Res Commun, 1999, 262(1): 223–230.
Leatherman J L, Kaestner K H, Jongens T A. Identification of a Mouse Germ Cell-Less Homologue with Conserved Activity in Drosophila [J]. Mech Dev, 2000, 92(2): 145–153.
Scholz S, Domaschke H, Kanamori A, et al. Germ Cell-Less Expression in Medaka (Oryzias latipes) [J]. Mol Reprod Dev, 2004, 67(1): 15–18.
Jongens T A, Ackerman L D, Swedlow J R, et al. Germ Cell-Less Encodes A Cell Type-Specific Nuclear Pore-Associated Protein and Functions Early in the Germ-Cell Specification Pathway of Drosophila [J]. Genes Dev, 1994, 8(18): 2123–2136.
Robertson S E, Dockendorff T C, Leatherman J L, et al. Germ Cell-Less Is Required Only During the Establishment of The Germ Cell Lineage of Drosophila and Has Activities Which Are Dependent and Independent of Its Localization to the Nuclear Envelope [J]. Dev Biol, 1999, 215(2): 288–297.
de La Luna S, Allen K E, Mason S L, et al. Integration of a Growth-Suppressing BTB/POZ Domain Protein with the DP Component of The E2F Transcription Factor [J]. Embo J, 1999, 18(1): 212–228.
Leatherman J L, Levin L, Boero J, et al. Germ Cell-Less Acts to Repress Transcription During the Establishment of the Drosophila Germ Cell Lineage [J]. Curr Biol, 2002, 12(19): 1681–1685.
Bardwell V J, Treisman R. The POZ Domain: A Conserved Protein-Protein Interaction Motif [J]. Genes Dev, 1994, 8(14):1664–1677.
Albagli O. The BTB/POZ Domain: A New Protein-Protein Interaction Motif Common to DNA-and Actin-Binding Proteins [J]. Cell Growth Differ, 1995, 6(9): 1193–1198.
Westerfield M. The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Brachydanio rerio)[M]. Eugene: University of Oregon Press, 1994.
Kimmel C B, Ballard W W, Kimmel S R, et al. Stages of Embryonic Development of the Zebrafish [J]. Dev Dyn, 1995, 203(3): 253–310.
Studier F W. A Strategy for High-Volume Sequencing of Cosmid DNAs: Random and Directed Priming with a Library of Oligonucleotides [J]. Proc Natl Acad Sci USA, 1989, 86(18): 6917–6921.
Li X, Peng H, Schultz D C, et al. Structure-Function Studies of The BTB/POZ Transcriptional Repression Domain from the Promyelocytic Leukemia Zinc Finger Oncoprotein [J]. Cancer Res, 1999, 59(20): 5275–5282.
Li W, Deng F, Wang H et al. Germ Cell-Less Expression in Zebrafish Embryos [J]. Development, Growrh and Differention, 2006, 48(5): 333–338.
Author information
Authors and Affiliations
Corresponding author
Additional information
Foundation item: Supported by the National Natural Science Foundation of China (30570968, 30370744)
Biography: DENG Fengjiao(1954–), female, Professor, research direction: developmental biology.
Rights and permissions
About this article
Cite this article
Deng, F., Cheng, P., Lan, Q. et al. Cloning and expression of BTB/POZ-domain from zebrafish gcl (germ cell-less), and appraisement of its polyclonal-antibody. Wuhan Univ. J. Nat. Sci. 13, 123–128 (2008). https://doi.org/10.1007/s11859-008-0124-2
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11859-008-0124-2