首页|大白菜BrMLP328的克隆、表达及功能验证

大白菜BrMLP328的克隆、表达及功能验证

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
  • 维普
[目的]克隆大白菜BrMLP328 基因,对其表达模式进行分析,并验证对花期调控的功能,为进一步探究该基因在大白菜成花调控过程的作用机理奠定基础.[方法]运用RT-PCR克隆BrMLP328,并进行生物信息学分析;利用RT-qPCR测定该基因的相对表达量;构建过表达载体并通过蘸花法转化野生型拟南芥,比较T2 代植株与野生型的开花时间差异.[结果]BrMLP328 的CDS全长为 456 bp,编码 151 个氨基酸,蛋白相对分子质量为 17 493.82 Da,定位于细胞核.BrMLP328 在大白菜茎中的表达量最高,根中次之,花蕾中最低;茎尖生长点中的表达量表现为春化后升高,之后在花芽分化阶段迅速下降,并维持在很低的水平.过表达BrMLP328 的拟南芥开花时间比野生型延迟了 1.46-3.09 d.[结论]从大白菜中克隆得到BrMLP328 基因,其表达量在不同组织及不同成花阶段有所不同,该基因能够延迟开花.
Cloning,Expression and Functional Identification of BrMLP328 Gene in Brassica rapa subsp.pekinensis
[Objective]The BrMLP328 gene of Chinese cabbage(Brassica rapa subsp.pekinensis)was cloned,its expression pattern was analysed,and its function in regulating flowering was verified.This may lay the basis for further investigation into the mechanism of the role of this gene in regulating flower formation in Chinese cabbage.[Method]BrMLP328 was cloned by RT-PCR and analysed by bioinformatics.RT-qPCR was used to determine the relative expression of the BrMLP328.An overexpression vector was constructed and transformed into wild-type Arabidopsis thaliana by flower-dipping method,and the difference in flowering time between the T2 generation plants and the wild type was compared.[Result]The coding sequence(CDS)of BrMLP328 has a total length of 456 bp,encoding 151 amino acids.The relative molecular weight of the protein was 17 493.82 Da and was localized in the nucleus.The expression of BrMLP328 was the highest in the stems of Chinese cabbage,followed by roots and lowest in flower buds.The expression in the growing point of the stem tip showed an elevated level after vernalization,after which it declined rapidly at the stage of flower bud differentiation and was maintained at a very low level.Compared with the wild type,the flowering time of the BrMLP328 of T2 generation of transgenic plants delayed by 1.46-3.09 d.[Conclusion]BrMLP328 was cloned from Chinese cabbage.The expression of BrMLP328 was different in different tissues and different flowering stages,and the gene could delay flowering.

Brassica rapa subsp.pekinensisBrMLP328flowering transformationgene cloningfunctional verification

杜泽光、任少文、张凤勤、李梅兰、李改珍、齐仙惠

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山西农业大学园艺学院,太原 030031

大白菜 BrMLP328 成花转变 基因克隆 功能验证

山西省基础研究计划山西省重点研发计划山西农业大学生物育种工程项目山西农业大学生物育种工程项目国家大宗蔬菜产业技术体系建设项目太原综合试验站项目大白菜种质资源创新与利用山西省科技创新重点团队项目

20210302124244202202140601005YZGC114YZGC121CARS-23-G092014131016

2024

10.13560/j.cnki.biotech.bull.1985.2023-1082

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(4)
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