Cloning and expression analysis of Atractylodes lancea AlWRKY2 gene
WRKY transcription factor plays a important role in material metabolism and various types of stress response during plant growth.In this study,based on the transcriptome data of Atractylodes lancea,the WRKY2 gene sequence was cloned and was named AlWRKY2(GenBank accession number:0R343916).Bioinformatics analysis was performed by software,and the results revealed that AlWRKY2 open reading frame was 975 bp,contained 324 amino acids.The encoded protein's theoretical isoelectric point was 6.62,and the molecular mass was 35 426.90.The secondary structure of AlWRKY2 was predict-ed to be mainly composed of random coils.Homologous alignment revealed that the amino acids encod-ed by WRKY2 genes of A.lancea,Quercus suber,Morella rubra,Theobroma cacao,Herrania umbratica,shared high sequence identity and had a typical WRKYGQK domain.The phylogenetic analysis indicated that the AlWRKY2 amino acid sequence of A.lancea was clustered in the same branch with Asteraceae plants and had high homology.The results of SDS-PAGE electrophoresis showed that the AlWRKY2 gene was successfully induced and expressed in Escherichia coli BL21(DE3)competent cells.The expected tar-get protein was successfully expressed as a soluble protein of about 50 kDa.The growth status of the re-combinant strain containing AlWRKY2 gene under NaCl and mannitol stress was significantly lower than that of the control strain and may have a negative regulatory effect.In different tissues of A.lancea,the re-sults of gene expression detected by qRT-PCR showed that the expression level of AlWRKY2 was the high-est in leaves and the lowest in stems.The expression of AlWRKY2 gene in A.lancea was increased under exogenous methyl jasmonate(MeJA)treatment,and the expression level was the highest at 24 h.The nu-clear localization analysis revealed that the AlWRKY2 gene resides within the nucleus.This study can pro-vide a reference for further study on the functions of WRKY transcription factors in A.lancea.
NETL
NSTL
万方数据
