Isolation and expression analysis of RgMed6 from Rehmannia glutinosa
10.7501/j.issn.0253-2670.2015.19.017
- Author:
Feng-Qing WANG
1
Author Information
1. College of Agronomy, Henan Agricultural University
- Publication Type:Journal Article
- Keywords:
Cloning;
Expression analysis;
Mediator subunit;
Rehmannia glutinosa L.;
RgMed6
- From:
Chinese Traditional and Herbal Drugs
2015;46(19):2919-2924
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone RgMed6 gene, which coded a subunit of mediator complex, from Rehmannia glutinosa, and to analyze the characteristics of protein sequence and gene expression. Methods: The transcriptional EST database of R. glutinosa was used to search analogs of AtMed6 gene by BLAST, the full-length open reading frame (ORF) of RgMed6 was obtained by assembling the ESTs. BLASTp, the online analysis tool of NCBI was used to get the homologous sequences of RgMed6, and MAFFT has been performed to analyze the multiple sequence alignment. Phylogenetic tree has been constructed using MEGA 6.0 software. Quantitative RT-PCR has been applied to detecting the transcription level of RgMed6 in five tissues as well as in tuberous roots or leaves under three stresses. Results: The cDNA sequence of RgMed6 containing 924 bp was obtained. The ORF of RgMed6 was 771 bp encoding 256 amino acids, which had typical structural domains and a potential nuclear localization signal (NLS). RgMed6 showed the highest expression level in leaves, followed by buds, but very weak in stems. The transcription level of RgMed6 mRNA was reduced under continuous cropping conditions in tuberous roots while it increased under salinity stress in leaves. Conclusion: RgMed6, a mediator subunit gene from R. glutinosa has been obtained for the first time, which can lay the foundation for further studies about its molecular function in development and responses to stress.
