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Cloning, silencing, and prokaryotic expression of strawberry sucrose synthase gene FaSus1

文献类型： 外文期刊

作者： Hua, L. N. 1 ; Zang, M. 1 ; Wang, S. F. 1 ; Li, Y. Z. 2 ; Shen, Y. Y. 1 ; Guo, J. X. 1 ;

作者机构： 1.Beijing Agr Univ, Coll Plant Sci & Technol, Beijing Key Lab New Technol Agr Applicat, Beijing, Peoples R China

2.Chinese Acad Agr Sci, Inst Agr Environm & Sustainable Dev, Water Resources & Dryland Farming Lab, Beijing, Peoples R China

关键词： Strawberry fruit ripening;hairpin-mediated RNA interference;sucrose synthase FaSus1;prokaryotic expression;enzymatic activity;sucrose

期刊名称：JOURNAL OF HORTICULTURAL SCIENCE & BIOTECHNOLOGY （ 影响因子：1.641； 五年影响因子：1.616 ）

ISSN：

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收录情况： SCI

摘要： The sucrose synthase (Sus) gene is regarded to be involved in strawberry fruit ripening; however, direct molecular and biochemical evidence is lacking. Here, the coding cDNA sequence of FaSus1 was cloned by RT-PCR and both a hairpin-mediated RNA interference pBI121 vector and a recombinant E. coli expression PET28a vector were constructed. These vectors were then used to transform strawberry fruit and for prokaryotic expression in strain BL21, respectively. The results showed that hairpin-mediated RNA silencing of FaSus1 led to a decrease in sucrose content and inhibited fruit ripening. Enzymatic activity analysis of FaSus1 showed that the reaction system contains 0.309 mg of recombinant FaSus1 protein, reaching 0.617 mg/mL. The cleavage activity of the enzyme was 0.25 U with a specific activity of 0.812 U/mg, whilst the synthetic activity of the enzyme was 0.057 U with a specific activity of 0.185 U/mg. This study provides physiological, molecular, and biochemical evidence to demonstrate that the FaSus1 has high sucrolysis activity and plays an important role in the regulation of strawberry fruit ripening.

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