Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.599793
Title: Isolation, characterisation and expression of genes encoding plant high mobility group HMG-I/Y proteins
Author: Gupta, R.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 1997
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Abstract:
The aim of the present study was to isolate gene(s) encoding HMG-I/Y proteins from higher plants and study the regulation of their expression. In order to isolate an HMG-I/Y gene a pea genomic library was screened with a pea HMG-I/Y cDNA resulting in the isolation of three different chimeric clones containing only the 3' end of the HMG-I/Y gene. The coding region (with a single intron of 201 bp) of the pea HMG-I/Y gene was isolated as a 795 bp fragment by PCR on genomic DNA. Several attempts using different approaches to isolate the promoter region of the pea HMG-I/Y gene were unsuccessful. The MHG-I/Y gene was present in a single-copy and expressed in all organs as determined by Southern and northern blot analysis. A cDNA encoding the HMG-I/Y protein from Arabidopsis thaliana was sequenced and characterised. The 903 bp cDNA encodes an HMG-I/Y protein of 204 amino acid residues with a calculated molecular mass of 22 kDa containing 4 copies of the 'AT-hook' motif. Southern blot analysis on Arabidopsis genomic DNA revealed the presence of a single copy of the HMG-I/Y gene. The HMG-I/Y gene was located near the top of chromosome 1 by RFLP analysis of F8 recombinant inbred lines. Northern blot analysis demonstrated the expression of the HMG-I/Y gene in all organs of Arabidopsis examined with the highest expression in flowers and developing siliques. A genomic clone encoding the HMG-I/Y protein was isolated from an Arabidopsis genomic library using the HMG-I/Y cDNA as a probe. A 6.3 kb genomic fragment was sequenced and includes the coding region (with a single 73 bp intron) of the HMG-I/Y gene and 5209 bp of 5' upstream sequence and 412 bp of 3' downstream sequence.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.599793  DOI: Not available
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