Determination of the relative expression levels of rubisco small subunit genes in Arabidopsis by rapid amplification of cDNA ends.
Multigene families are common in higher organisms. However, due to the close similarities between members, it is often difficult to assess the individual contribution of each gene to the overall expression of the family. In Arabidopsis thaliana, there are four genes encoding the small subunits (SSU) of ribulose-1.5-bisphosphate carboxylase oxygenase (rubisco) whose nucleotide sequences are up to 98.4% identical. In order to overcome the technical limitations associated with gene-specific probes (or primers) commonly used in existing methods, we developed a new gene expression assay based on the RACE (rapid amplification of cDNA ends) technique with a single pair of primers. With this RACE gene expression assay, we were able to determine the relative transcript levels between four Arabidopsis SSU genes. We found that the relative SSU gene expression differed significantly between plants grown at different temperatures. Our observation raises the possibility that an adaptation of rubisco to the environment may be achieved through the specific synthesis of the SSU proteins, which is determined by the relative expression levels between the SSU genes.
Yoon
M
M
School of Biological Sciences, University of Auckland, Auckland, New Zealand. m.yoon@auckland.ac.nz
Putterill
J J
JJ
Ross
G S
GS
Laing
W A
WA
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D001345:Autoradiography; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018749:RNA, Plant / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics*; D012680:Sensitivity and Specificity; D013696:Temperature
D018076:DNA, Complementary; D018749:RNA, Plant; D012273:Ribulose-Bisphosphate Carboxylase