Cloning, prokaryotic expression, and bioinformatics analysis of <i>PaPR10-1</i> gene from <i>Picea asperata</i> Mast.

Li Cheng-Song; Liu Li-Juan; Liang Fang; Zhao Wei-Dong; Yang Chun-Lin; Liu Ying-Gao

doi:10.11913/PSJ.2095-0837.22140

Plant Science Journal > 2023 > 41(2): 224-233. > DOI: 10.11913/PSJ.2095-0837.22140 CSTR: 32231.14.PSJ.2095-0837.22140

Li CS，Liu LJ，Liang F，Zhao WD，Yang CL，Liu YG. Cloning, prokaryotic expression, and bioinformatics analysis of PaPR10-1 gene from Picea asperata Mast.[J]. Plant Science Journal，2023，41（2）：224−233. DOI: 10.11913/PSJ.2095-0837.22140

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Cloning, prokaryotic expression, and bioinformatics analysis of PaPR10-1 gene from Picea asperata Mast.

National Forestry and Grassland Administration Key Laboratory of Forest Resources Conservation and Ecological Safety on the Upper Reaches of the Yangtze River, Sichuan Agricultural University, Chengdu 611130, China

Funds: This work was supported by a grant from the Key Project of Education Department of Sichuan (09ZA068).

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Received Date: August 29, 2022
Revised Date: October 29, 2022
Available Online: May 05, 2023

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Abstract

Abstract

PR10 is an important member of the pathogenesis-related protein (PR) family, which enhances the ability of plants to resist external stresses. In this study, a PR10 gene from Picea asperata Mast. PaPR10-1 was obtained by RT-PCR, with sequence characteristics then analyzed and prokaryotic expression system further constructed and optimized. Ribonuclease activity was then analyzed using the substrate method in vitro. Results showed that the open reading frame (ORF) of PaPR10-1 was 486 bp in length and encoded a protein with a 161 amino acid. Without a transmembrane structure and signal peptide, the PaPR10-1 protein was an intracellular protein with two conserved domains, “P-Loop” and Bet-v1-like, respectively. Phylogenetic analysis showed that PaPR10-1 was closely related to the PR10 protein of Pinus pinaster Aiton. Optimal expression conditions for the gene were 0.2 mmol/L IPTG at 30℃ for 1 h induction, and the expressed target protein exhibited ribonuclease activity.
- Picea asperata,
- PR10 gene,
- Gene cloning,
- Bioinformatics,
- Prokaryotic expression

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References

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Cloning, prokaryotic expression, and bioinformatics analysis of PaPR10-1 gene from Picea asperata Mast.