Reverse transcription quantitative-PCR of three genes with high homology encoding 3-hydroxy-methylglutaryl-CoA reductase in rice.
Reverse transcription followed by RT Q-PCR is useful for the systematic measurement of changes in gene expression. RT Q-PCR with two pairs of primers for each gene was used for relative expression of three genes with high homology encoding 3-hydroxy-methylglutaryl-CoA reductase (HMGR) in rice. At various growth stages of etiolated seedling and various times after UV-irradiation treatment, RT Q-PCR of each HMGR gene showed a consistent pattern of relative expression with the RT Q-PCR data, using two pairs of primers, giving a high degree of accuracy. Furthermore, the different expression levels of three HMGR genes in a sample were determined by diluting the cDNA concentration. These results indicate that RT Q-PCR with only one pair of primers for a gene can quantify the relative expression and that the high expression level of HMGR2 could be quantified in comparison to the low level of HMGR1 expression.
School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea.
Bo-Ra
BR
Kim
Hee-Young
HY
Nam
Soo-Un
SU
Kim
Tongkun
T
Pai
Yung-Jin
YJ
Chang
D020869:Gene Expression Profiling / Q000379:methods*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012275:Oryza / Q000201:enzymology* / Q000235:genetics*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods; D017386:Sequence Homology, Amino Acid
D006903:Hydroxymethylglutaryl CoA Reductases