Full-length cDNA cloning of AsNINJA1 gene in Aquilaria sinensis and MeJA-induced expression in callus
10.7501/j.issn.0253-2670.2014.20.019
- Author:
Yong-Cui LIAO
1
Author Information
1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College
- Publication Type:Journal Article
- Keywords:
Aquilaria sinensis (Lour.) Gilg;
AsNINJA1 gene;
Expression analysis;
MeJA;
Rapid amplification of cDNA ends;
Sequence analysis
- From:
Chinese Traditional and Herbal Drugs
2014;45(20):2968-2973
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone the full-length cDNA of interacting protein of JAZ (NINJA) gene in Aquilaria sinensis, and to provide the basic information for further study on gene function in sesquiterpenes biosynthesis pathway. Methods: With the total RNA as template, the full-length cDNA of NINJA in A. sinensis was cloned through RACE technique and RT-PCR method. The bioinformatics of cloing NINJA gene was analyzed as well. The expression mode of this gene was with MeJA treatment in A. sinensis callus detected by qRT-PCR method. Results: The full-length cDNA (1 982 bp) of NINJA gene in A. sinensis, named as AsNINJA1 was obtained with an open reading frame of 1 221 bp and encoding 406 amino acids. The relative molecular mass of AsNINJA1 protein calculated was 43 697, and the isoelectric point was 6.02. The qRT-PCR results indicated that MeJA treatment could stimulate the increase of mRNA expression of AsNINJA1; There was a sharp rise at 4 h with nearly 100 times higher than the control (without MeJA treatment), then dropped significantly. Conclusion: The full-length cDNA sequence of AsNINJA1 gene is obtained; AsNINJA1 is extremely sensitive to MeJA treatment, and responded to the early damage.
