Analysis of the Gene Structure,Prokaryotic Expression and Expression Pattern of StRTG2 in Setosphaeria turcica

doi:10.7668/hbnxb.20192146

Abstract

Abstract: In order to clarify the function of the StRTG2 gene of Setosphaeria turcica and its expression pattern in different developmental stages of the pathogen, the amino acid sequence encoded by the yeast ScRTG2 gene was used for BlastP, and the homologous gene sequence in Setosphaeria turcica was obtained, and named it as StRTG2;The whole genome DNA and cDNA of wild type 01-23 of Setosphaeria turcica were used respectively as a template to clone the gene;Use bioinformatics technology to analyze the physicochemical properties, domain prediction, and subcellular location prediction of StRtg2;Use MEGA 7.0 to analyze the evolutionary relationship of Rtg2;Amplify the ORF sequence of StRTG2 to construct a prokaryotic expression vector of the gene. After transforming into E.coli BL21, IPTG was used to induce express;Finally, the RNA-Seq database was used to analyze the main growth and development stages of the StRTG2 gene (hyphae, conidia, germ tubes, appressorium and invading nails). The main results were as follows:The gene was cloned and found to be 1 695 bp in length without introns;the protein encoded by the gene was composed of 564 amino acids, with a theoretical isoelectric point (pI) of 6.96, and had a typical Ppx-Gppa conserved domain. The cell location prediction results showed that the protein was distributed in all parts of the cell, and the possibility of being distributed in the cytoplasm was the greatest;The analysis of the evolutionary relationship of the protein showed that the StRtg2 protein had the highest homology with the homologous protein of Stemphylium lycopersici, followed by Saccharomyces cerevisiae;Prokaryotic expression of the gene, SDS-PAGE gel showed that the size of the protein was 62 ku, which was in line with the expected size;Analysis of the expression pattern revealed that the gene was expressed in the 5 stages of pathogen development, among them, the expression of the germ tube and the invading nail period was significantly reduced. This research can lay the foundation for further analysis of the function of StRTG2 gene.

Key words: Setosphaeria turcica, Gene cloning, StRTG2 gene, Prokaryotic expression, Expression pattern analysis

CLC Number:

Q78
S513.03

LI Moxiao, BIAN Zhe, ZHOU Qihui, LIU Yuwei, GONG Xiaodong, GU Shouqin, HAN Jianmin. Analysis of the Gene Structure,Prokaryotic Expression and Expression Pattern of StRTG2 in Setosphaeria turcica[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 191-196. doi: 10.7668/hbnxb.20192146.

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