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西瓜ClPP2C3克隆及表达分析

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[目的]蛋白磷酸酶 2C(protein phosphatase 2C,PP2C)是动植物中均存在的一类蛋白磷酸酶,拟探究其在西瓜果实成熟过程中发挥的重要作用.[方法]通过逆转录PCR(reverse transcription-polymerase chain reaction,RT-PCR)从栽培类型西瓜'97103'中克隆ClPP2C3,并对其进行生物信息学、表达模式和亚细胞定位分析.[结果]西瓜ClPP2C3 的cDNA序列长度为 1 317 bp,编码 438 个氨基酸,其分子量大小为 47.81 kD,蛋白质等电点为 5.12.ClPP2C3 包含 1 个PP2C保守结构域,与负调控果实成熟的番茄SlPP2C3、草莓FaABI1 具有较高的同源性.西瓜ClPP2C3在细胞核表达.ClPP2C3在含糖量高的栽培品种中表达量显著高于含糖量低的野生品种.栽培品种ClPP2C3的 2 kb片段长度启动子活性显著高于野生品种,而 1 kb片段长度启动子活性之间无显著差异.[结论]由 1-2 kb区间SNP导致的启动子活性差异对不同含糖量品种ClPP2C3表达量存在影响.
Cloning and Expression Analysis of ClPP2C3 in Citrullus lanatus
[Objective]Protein phosphatase(PP2C)is a type of protein phosphatase that exists in both animals and plants,In order to explore its important role in the ripening process of watermelon fruits.[Method]In this study,a PP2C gene ClPP2C3 was cloned in watermelon(Citrullus lanatus)by reverse transcription-polymerase chain reaction(RT-PCR),and its bioinformatics,expression pattern and subcellular localization analysis were conducted.[Result]The cDNA sequence length of ClPP2C3 in watermelon was 1 317 bp and it encoded 438 amino acids.The molecular weight of ClPP2C3 protein was 47.81 kD and its isoelectric point was 5.12.The ClPP2C3 protein contained one PP2C conserved domain and it had high homology with tomato SlPP2C3 in tomato and FaABI1 in strawberry that negatively regulated fruit ripening.ClPP2C3 in the watermelon was located in the nucleus.The gene quantitative expression analysis showed that the expression of ClPP2C3 in high-sugar-containing watermelons was significantly higher than that in low-sugar-containing watermelons.The 2 kb promoter(up 2 kb to ATG)activity of cultivated watermelon was significantly higher than that in ancestral watermelons,while there was no significant difference in the 1 kb promoter(up 1 kb to ATG)activity detected between these two species.[Conclusion]The SNPs in 1-2 kb promoter region might result in the difference of promoter activity in different varieties.

Citrullus lanatusClPP2C3gene cloningsubcellular localizationanalysis of promoter activity differences

朱毅、柳唐镜、宫国义、张洁、王晋芳、张海英

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中国农业大学园艺学院,北京 100193

北京市农林科学院蔬菜研究所,北京 100097

广西壮族自治区农业科学院园艺研究所,南宁 530007

西瓜 ClPP2C3 基因克隆 亚细胞定位 启动子活性差异分析

北京市农林科学院蔬菜研究所改革与发展项目北京市农林科学院青年基金项目国家西甜瓜产业技术体系项目北京市特色作物创新团队项目广西壮族自治区八桂学者项目

KYCX202301QNJJ202206CARS-25BAIC04-20232016A11

2024

10.13560/j.cnki.biotech.bull.1985.2023-0709

生物技术通报
中国农业科学院农业信息研究所

生物技术通报

CSTPCD北大核心
影响因子：0.505
ISSN：1002-5464
年,卷(期)：2024.40(1)
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