Cloning of a gene encoding an antigen associated with the centrosome in Drosophila
Abstract
The monoclonal antibody Bx63 recognizes a centrosomal antigen of Drosophila melanogaster by indirect immunofluorescence and identifies two proteins, with apparent molecular weights of 185 x 10³ and 66 x 10³, on Western blots. We have used this antibody to isolate five clones (λcs1, -2, -3, -4 and λj63) from λgt11 expression libraries of Drosophila DNA. Using polyclonal anti-centrosomal sera raised against both immunoaffinity-purified Bx63 antigen and electrophoretically purified fusion protein from clone λcs3, we have demonstrated that the fusion proteins encoded by four of these clones (λcs1-4) share at least two epitopes with the 185 x 10³ M_r centrosomal antigen. This indicates that clones λcs1-4 contain DNA from the gene coding for this protein. The four clones are independent isolates from a single chromosomal site, which we show by in situ hybridization to correspond with salivary gland chromosome region 88E 4-8. A low-abundance transcript of approximately 4.0 x 10³ bases corresponding to the cloned gene is detected in all stages of the Drosophila life-cycle.
Additional Information
© 1988 by Company of Biologists. (Received 20 October 1987 - Accepted 9 December 1987) This work was supported by a grant from the Cancer Research Campaign (CRC) for which we are most grateful. D.M.G. holds a Career Development Award from the CRC and received short term fellowships from EMBO that enabled him to visit Tübingen in July 1984 and May 1985. S.E.M. held an MRC Research Studentship. We would like to thank Claudio Sunkel for carrying out the in situ hybridization experiment shown in Fig. 9, and Nicola Williamson for assistance with the early stages of this work. Herbert Jäckle and Reinhard Schuh introduced us to the techniques of screening λgt11 libraries with antibodies and M. Wilcox and G. Pflugfelder provided libraries.Attached Files
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- 105700
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- CaltechAUTHORS:20200930-144645910
- Cancer Research Campaign
- European Molecular Biology Organization (EMBO)
- Medical Research Council (UK)
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