A Rab-related small GTP binding protein is predominantly expressed in root nodules of Medicago sativa.
Rab-related small GTP-binding proteins are known to be involved in the regulation of the vesicular transport system in eucaryotic cells. In this paper we report the isolation of the cDNA clone MS- rab11f from Medicago sativa (alfalfa) root nodules using a combination of RT-PCR and SSCP analysis. MS- rab11f shows high homology to the Rab-related cDNA clone LJ- rab11f from Lotus japonicus root nodules. The MS-Rab11F protein expressed in Escherichia coli was found to bind GTP, confirming that the isolated cDNA indeed codes for a small GTP-binding protein. Expression analysis by RT-PCR demonstrated that MS- rab11f is preferentially expressed in root nodules of alfalfa. Using the cDNA-sequence of MS-rab11f, a peptide-specific antibody was generated. Western blot analysis with this antibody revealed that two Rab11F isoforms, designated MS-Rab11FA and MS-Rab11FB, are found in M. sativa root nodules.
Schiene
K
K
Biologie-Genetik, Universität Bielefeld, Postfach 100 131, 33501 Bielefeld, Germany.
Donath
S
S
Brecht
M
M
Pühler
A
A
Niehaus
K
K
D000595:Amino Acid Sequence; D015153:Blotting, Western; D006160:Guanosine Triphosphate / Q000378:metabolism; D000455:Medicago sativa / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006160:Guanosine Triphosphate; D020559:Monomeric GTP-Binding Proteins