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.
Kim
Bo-Ra
BR
School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea.
Nam
Hee-Young
HY
Kim
Soo-Un
SU
Pai
Tongkun
T
Chang
Yung-Jin
YJ
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