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[Cloning and Expression Analysis of Acetyl-CoA C-acetyltransferase Gene in Isodon rubescens]
Objective:
To clone the acetyl-CoA C-acetyl transferase( AACT) gene from Isodon rubescens, and to analyze the bioinformatics and expression of the gene.
Methods:
According to the IrAACT gene sequence of Isodon rubescens transcriptome,a pair of primers was designed, and the ORF of cDNA sequence was obtained by reverse transcription PCR. Bioinformatic analysis of this gene and its corresponding protein were performed. Real-time quantitative PCR( q PCR) was used to detect the relative expression levels of IrAACT different tissues of Isodon rubescens.
Results:
The IrAACT cDNA sequence contained a 1 254 bp open reading frame and encoded a predicted protein of 417 amino acids. IrAACT had extensive homology with AACTs from other plant species, such as Salvia miltiorrhiza, et al. Bioinformatic analysis showed that IrAACT-encoding protein contained the thiolase Ⅱ catalytic domain. q PCR analysis showed that the expression of IrAACT was tissue-specific, and accumulation of transcripts was greater in flowers and leaves, followed by stems, roots and callus.
Conclusion:
It is the first time to report IrAACT gene and its relative expression level. The results will provide a groundwork for studying the function of IrAACT in terpenoid biosynthesis of Isodon rubescens.
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