Acta Agriculturae Boreali-Sinica ›› 2022, Vol. 37 ›› Issue (5): 36-44. doi: 10.7668/hbnxb.20193096

Special Issue: Plant protection Biotechnology

• Crop Genetics & Breeding·Germplasm Resources·Biotechnology • Previous Articles     Next Articles

Cloning and Prokaryotic Expression of ACS Gene from Paeonia lactiflora Pall.

XIAO Shikui1, LI Fang1, ZHANG Wenting2, LÜ Shufang1, SHI Guoan1, WU Jiang3, , FAN Bingyou1,   

  1. 1 College of Agriculture,Henan University of Science and Technology,Luoyang 471023,China
    2 SINOMACH TDI International Engineering Co., Ltd, Luoyang 471003, China
    3 College of Information Engineering,Yulin University,Yulin 719000,China
  • Received:2022-06-21 Published:2022-10-28
  • Contact: WU Jiang, FAN Bingyou


In order to explore the function of ACS gene in herbaceous peony,a full-length cDNA sequence of PlACS cDNA in Paeonia lactiflora was obtained,RACE technique and bioinformatic methods were used to analyze the protein sequence which it encoded.The CDS of PlACS was subcloned,the prokaryotic expression vector of PlACS was constructed based on pET32a vector,and then the highly efficient prokaryotic expression system was established.The results showed that the total length of PlACS cDNA(GenBank accession JX512359)was 1 752 bp,which encoded 492 amino acids.Seven conserved regions and active sites K278 were detected in PlACS protein.Phylogenetic tree analysis showed that PlACS was highest homological with ACS of P.suffruticosa.PlACS protein was determined structurally to be 40.04% α-helix,16.26% β-extended strand,6.91% β-turn and 36.79% random coil.Protein 3D structure homology modeling predicted that PlACS existed as homodimers.The optimal expression condition of PlACS protein was that when the cell density of genetic engineering strain A600 reached 0.2,IPTG with a final concentration of 0.1 mmol/L was added,and the recombinant protein was expressed for two hours at 37 ℃.It was of great significance to acquire PlACS recombinant protein with biological activity by denaturation & renaturation and identify its enzymatic activity in vitro.

Key words: Paeonia lactiflora, PlACS, Gene cloning, Sequence analysis, Prokaryotic expression

Cite this article

XIAO Shikui, LI Fang, ZHANG Wenting, LÜ Shufang, SHI Guoan, WU Jiang, FAN Bingyou. Cloning and Prokaryotic Expression of ACS Gene from Paeonia lactiflora Pall.[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 36-44. doi: 10.7668/hbnxb.20193096.

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