Preparation and Function Verification of Recombinant Protein of Chalcone Isomerase Gene from <i>Rhododendron Delavayi</i>

WANG Yuhan; SUN Shiyu; JU Zhigang; SUN Wei; XU Xiaorong

doi:10.19303/j.issn.1008-0384.2023.05.009

Volume 38 Issue 5

May 2023

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Article Navigation > Fujian Journal of Agricultural Sciences > 2023 > 38(5): 574-582

WANG Y H, SUN S Y, JU Z G, et al. Preparation and Function Verification of Recombinant Protein of Chalcone Isomerase Gene from Rhododendron Delavayi [J]. Fujian Journal of Agricultural Sciences，2023，38（5）：574−582 doi: 10.19303/j.issn.1008-0384.2023.05.009

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WANG Y H, SUN S Y, JU Z G, et al. Preparation and Function Verification of Recombinant Protein of Chalcone Isomerase Gene from Rhododendron Delavayi [J]. Fujian Journal of Agricultural Sciences，2023，38（5）：574−582 doi: 10.19303/j.issn.1008-0384.2023.05.009

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Preparation and Function Verification of Recombinant Protein of Chalcone Isomerase Gene from Rhododendron Delavayi

doi: 10.19303/j.issn.1008-0384.2023.05.009

WANG Yuhan^1
,,
SUN Shiyu¹,
JU Zhigang²,
SUN Wei^{1, 3
,
,},
XU Xiaorong^{1, 3
,
,}

1.
Key Laboratory of Plant Physiology and Development Regulation/School of Life Science, Guizhou Normal University, Guiyang, Guizhou　550025, China
2.
Pharmacy College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou　550025, China
3.
Key Laboratory of State Forestry Administration on Biodiversity Conservation in Karst Mountain Area of Southwest of China, Guiyang, Guizhou　550025, China

Received Date: 2022-09-14
Accepted Date: 2022-09-14
Rev Recd Date: 2023-03-10

Available Online: 2023-05-24

Publish Date: 2023-05-28

Abstract

Abstract

Objective Recombinant protein of chalcone isomerase (CHI) gene in Rhododendron delavayi was prepared and its activity was verified. This will provide theoretical basis for analyzing the function of CHI, and also lays a foundation for improving flower color and increasing medicinal ingredientst. Method Primers were designed based on the sequence of RdCHI1, and the prokaryotic expression vector constructed. Conditions to induce the expression of soluble recombinant protein were optimized. Activity of the prepared protein was verified by an in vitro enzymatic assay. Result The successfully constructed RdCHI1 prokaryotic expression vector was expressed in the supernatant under the optimized induction that applied 0.35 mmol·L⁻¹ IPTG at 15 ℃ for 36 h. The nickel column-purified recombinant protein significantly hastened the conversion of naringin chalcone to naringin in an in vitro assay. Conclusion As a type I CHI, RdCHI1 significantly accelerated the convertion from naringin chalcone to naringin and increase the accumulation od flavonoids.
- Rhododendron delavayi,
- chalcone isomerase,
- prokaryotic expression vector,
- soluble recombinant protein,
- enzyme assay

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References(48)

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