Molecular and biochemical characterisation of sucrose and amino acid carriers in Ricinus communis
Molecular and biochemical characterisation of sucrose and amino acid carriers in Ricinus communis
Using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) two putative amino acid carrier partial-length clones, RAAC1 and RAAC2, were obtained from Ricinus root RNA. These clones were shown to be highly homologous with the AAP family of amino acid carriers from Arabidopsis. Northern analyses of RAAC1 and RAAC2, and a partial-length sucrose carrier clone, RSC1, have identified the expression of all three carriers in the seedling and leaf tissues. The strongest expression of RAAC1, RAAC2 and RSC1 was evident in the cotyledons, reflecting the importance of this organ in the absorption of solutes from the endosperm during germination. Using in situ hybridisation, the expression of the RAAC1 and RAAC2 amino acid carriers was localised to the root stele, in particular within the cells immediately adjacent to the xylem poles. It is proposed that these cells represent the pericycle and/or xylem parenchyma, and the expression of amino acid carriers in these cells may indicate a role xylem loading or possibly in the initiation of lateral root development.
Three yeast strains were engineered for the future isolation of Ricinus sucrose and amino acid carriers by functional complementation. Two strains, 2512C ura3-52 and JB2, have been constructed for the isolation and functional expression of amino acid carriers. The third strain, SUSY7 ura3-52, has been designed for the isolation of plant sucrose carrier genes. Transformation of this yeast with an Arabidopsis cDNA library resulted in the isolation of the clone, A1, which functionally complements the growth of this yeast on sucrose. Molecular characterisation of the A1 clone suggested that it is unrelated to other higher plant sucrose carriers.
University of Southampton
1996
Bick, Julie-Ann
(1996)
Molecular and biochemical characterisation of sucrose and amino acid carriers in Ricinus communis.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) two putative amino acid carrier partial-length clones, RAAC1 and RAAC2, were obtained from Ricinus root RNA. These clones were shown to be highly homologous with the AAP family of amino acid carriers from Arabidopsis. Northern analyses of RAAC1 and RAAC2, and a partial-length sucrose carrier clone, RSC1, have identified the expression of all three carriers in the seedling and leaf tissues. The strongest expression of RAAC1, RAAC2 and RSC1 was evident in the cotyledons, reflecting the importance of this organ in the absorption of solutes from the endosperm during germination. Using in situ hybridisation, the expression of the RAAC1 and RAAC2 amino acid carriers was localised to the root stele, in particular within the cells immediately adjacent to the xylem poles. It is proposed that these cells represent the pericycle and/or xylem parenchyma, and the expression of amino acid carriers in these cells may indicate a role xylem loading or possibly in the initiation of lateral root development.
Three yeast strains were engineered for the future isolation of Ricinus sucrose and amino acid carriers by functional complementation. Two strains, 2512C ura3-52 and JB2, have been constructed for the isolation and functional expression of amino acid carriers. The third strain, SUSY7 ura3-52, has been designed for the isolation of plant sucrose carrier genes. Transformation of this yeast with an Arabidopsis cDNA library resulted in the isolation of the clone, A1, which functionally complements the growth of this yeast on sucrose. Molecular characterisation of the A1 clone suggested that it is unrelated to other higher plant sucrose carriers.
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Published date: 1996
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Local EPrints ID: 459932
URI: http://eprints.soton.ac.uk/id/eprint/459932
PURE UUID: ea7afd18-3a92-4074-ae82-2fdb73c04fd2
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Date deposited: 04 Jul 2022 17:28
Last modified: 04 Jul 2022 17:28
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Author:
Julie-Ann Bick
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