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| Cloning and Prokaryotic Expression of the UDP-glycosyltransferaseGene PpUGT2 from Paris polyphylla var. chinensis |
| WANG Li-Ya1, WANG Jing-Han2, ZHOU Hui-Min1, ZHANG Yao1, CHEN Wen-Qiang1, LU Zhi-Guo1, ZHU Li-Na1*, TANG Qi1* |
1 College of Horticulture, Hunan Agricultural University, Changsha 410128, China;
2 College of Landscape Architecture, Central SouthUniversity of Forestry and Technology, Changsha 410128, China |
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Abstract Saponins are important active components of Paris polyphylla chinensis, but their biosynthetic pathway has not been fully elucidated. Research suggests that UPD-glycosyltransferases (UGTs) are key enzymes in the downstream biosynthetic pathway of polyphyllin. This study based on the correlation analysisbetween the transcriptome of P. polyphylla var. chinensis and the total saponins content, a glycosyltransferasecandidate gene PpUGT2 (GenBank No. pq588702) was screened out, which might be involved in the biosynthesis of polyphyllin. The PpUGT2 gene was cloned using PCR technology, and an expression vector was constructed for prokaryotic expression. Expression was induced using IPTG, followed by purification ofthe recombinant protein. Concurrently, qPCR was employed to analyze the expression patterns of PpUGT2across different parts of P. polyphylla var. chinensis. The results indicated that PpUGT2 had a total length of1 773 bp, encoded a protein consisting of 590 amino acids with a relative molecular weight of 64.6 kD. This hydrophilic protein was unstable and exhibited good lipid solubility. SDS-PAGE analysis confirmed the expression of the PpUGT2 recombinant protein at approximately 64.6 kD. Furthermore, qPCR resultsdemonstrated expression of the PpUGT2 gene in roots, stems, leaves, and young fruits of P. polyphylla var.chinensis, with a distinct expression hierarchy of roots>stems>leaves>young fruits, suggesting a predominant role of this gene in root tissues. This study enriches the study of UGT in P. polyphylla var. chinensis and provides a theoretical basis for further analysis of the biosynthesis pathway of polyphyllin.
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Received: 19 September 2024
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Corresponding Authors:
*tangqi@hunau.edu.cn;zhulina_2021@163.com
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