线果芥CpNSP5基因的克隆及表达分析
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国家转基因生物新品种重大专项(2013ZX08005004)


Clonging and Expression Analysis of the CpNSP5 Gene in Conringia planisiliqua L.
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    摘要:

    以该实验室前期从线果芥(Conringia planisiliqua L.)中通过mRNA差异显示技术筛选到的414 bp干旱响应相关核心片段为基础,采用RACE技术对该片段进行全长克隆,序列比对分析发现,该片段与拟南芥的AtNSP5基因相似性较高,命名为CpNSP5。CpNSP5基因全长1 228 bp,开放阅读框966 bp,编码321个氨基酸。推测蛋白分子量为35.034 5 kD,等电点为5.41。蛋白二级结构预测包含26个β折叠,具有典型的Kelch repeat 结构。系统进化分析发现,CpNSP5与白菜亲缘关系最近。实时荧光定量PCR检测发现,CpNSP5基因在20% PEG6000和200 mmol·L-1 NaCl胁迫下均受到不同程度诱导,说明CpNSP5基因可能与线果芥响应逆境胁迫有关。该研究结果为作物的抗旱育种提供了候选基因。

    Abstract:

    Using the core fragment screened by mRNA differential display technology in our laboratory preliminary study, which is about 414 bp length deserved from the Conringia planisiliqua L. under drought stress, we cloned the full length cDNA by RACE technology and sequence analysis showed it has higher similarity to AtNSP5, so it was named CpNSP5. The length of CpNSP5 is about 1 228 bp and with an open reading frame of 966 bp. It encodes a protein composed of 321 amino acids whose molecular weight is 35.034 5 kD and the isoelectric point is 5.41. The secondary structure of the CpNSP5 protein contained 26 βsheets and had the Kelch repeat conserved domain. According to phylogentic tree analysis, CpNSP5 kept close relationship with Brassica rapa. Realtime quantitative PCR analysis showed that the transcript of CpNSP5 was induced strongly under 20% PEG6000 stress and 200 mmol·L-1 NaCl stress. These results indicated that CpNSP5 might be involved in stress responses of C. planisiliqua L. which can provide a candidate gene for crop drought stress breeding.

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朱燕飞,陈全家,姚正培,等.线果芥CpNSP5基因的克隆及表达分析[J].西北植物学报,2018,38(1):48-54

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  • 在线发布日期: 2018-02-05
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