cDNA cloning and analysis of a flg variant fibroblast growth factor receptor from Xenopus laevis

Chen, Gang (1993) cDNA cloning and analysis of a flg variant fibroblast growth factor receptor from Xenopus laevis. Masters thesis, Memorial University of Newfoundland.

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    Available under License - The author retains copyright ownership and moral rights in this thesis. Neither the thesis nor substantial extracts from it may be printed or otherwise reproduced without the author's permission.
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Abstract

1. A cDNA library was constructed with mRNA isolated from stage 8 Xenopus embryos. -- 2. A full length cDNA clone encoding a FGFR-1/flg gene was isolated by screening this cDNA library. It is designated XFGFR-A3. -- 3. The XFGFR-A3 clone was sequenced by dideoxynucleotide chain termination method on both strands with synthetic oligonucleotides. It is 3863 bp in length and is predicted to encode an 810 aa protein. -- 4. The XFGFR-A3 clone contains two dipeptide deletions, Val⁴²³⁻ Thr⁴²⁴ and Pro⁴⁴¹Ser⁴⁴², which are different from the published XFGFR-1 sequence ( Figure 8 ) . The Pro⁴⁴¹Ser⁴⁴² deletion has been described previously, however, this is the first report of the Val⁴²³⁻Thr⁴²⁴ deletion in Xenopus. Both dipeptide deletions result in the removal of consensus phosphorylation sites for Protein Kinase C ( PKC ) which may have consequences on intracellular signal transduction and the regulation of embryonic development. -- 5. RT-PCR results showed that XFGFR-A3 was expressed at all stages of Xenopus embryonic development. -- 6. The RNase protection experiment showed that XFGFR-A3 is a minor form of the XFGFR-1 in all stages of Xenopus embryonic development. Like the wild type XFGFR-1, XFGFR-A3 is also uniformly expressed throughout Xenopus development. -- 7. The XFGFR-A3 genomic DNA sequence covering the VT deletion region was sequenced. The VT deletion is located at an exon/intron boundary and comparison with the cDNA sequence suggests that the XFGFR-A3 variant arises from the use of an alternate 5'splice donor site. -- 8. Expression vectors containing an insert covering the VT deletion region fused to the cDNA encoding GST were constructed by using XFGFR-A2 and XFGFR-A3. A PKC assay using the purified fusion proteins products showed that Thr⁴²⁴ of XFGFR-A2 can be phosphorylated by PKC in vitro.

Item Type: Thesis (Masters)
URI: http://research.library.mun.ca/id/eprint/5622
Item ID: 5622
Additional Information: Bibliography: leaves 124-153.
Department(s): Medicine, Faculty of
Date: 1993
Date Type: Submission
Library of Congress Subject Heading: Fibroblast growth factors; Xenopus laevis; Developmental genetics; Mesoderm
Medical Subject Heading: Fibroblast Growth Factors; Genetics; Xenopus laevis; Mesoderm

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