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烟草NtCAD基因克隆与表达分析

Cloning and expression analysis of NtCAD gene from Nicotiana tabacum

  • 摘要: 为明确CAD基因在烟草中的功能和调控方式,通过同源克隆法从烟草K326中克隆得到1个NtCAD基因,其开放阅读框(Open reading frame,ORF)含有1074bp,编码357个氨基酸,预测分子量38.99 kDa、等电点5.54,具有CAD蛋白保守结构域,启动子区域含有参与低温和脱落酸应答的顺式作用元件。NtCAD在烟草花蕾、侧根和须根中的表达量相对较高,茎节、茎秆的表达量较低。低温(4℃)处理能显著诱导NtCAD表达,脱落酸(ABA)处理后NtCAD表达量先升高后下降又升高,说明该基因可能参与了抵御非生物胁迫。

     

    Abstract: In order to clarify the function and regulation patterns of CAD gene in tobacco, a NtCAD gene was isolated from tobacco cultivar K326(Nicotiana tabacum)by homologous cloning. This gene had an open reading frame(ORF)of 1 176 bp, encoding 391 amino acids, and its predicted molecular weight was 38.99 kDa with an isoelectric point of 5.54. The amino acid sequence of NtCAD had conserved domains of CAD protein. The expression level of NtCAD gene in the flower bud, lateral root and fibrous root was relatively high, but it was relatively low in the stalk node and stalk. A low temperature (4 ℃) treatment significantly induced the expression of NtCAD, while the treatment of abscisic acid(ABA)caused the expression of NtCAD to display a cyclical increase, followed by decrease and increase again, which indicated that NtCAD might be involved in the resistance to abiotic stress.

     

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