WANG Li-juan,GAO Wei-feng,GUAN Cui-ping,et al.Cloning and Expression Analysis of WRKY Gene Segment in Lycium barbarum L.[J].Northern Horticulture,2013,37(20):89-94.
枸杞WRKY基因片段克隆与表达分析
- Title:
- Cloning and Expression Analysis of WRKY Gene Segment in Lycium barbarum L.
- 文章编号:
- 1001-0009(2013)20-0089-06
- 分类号:
- Q 785
- 文献标志码:
- A
- 摘要:
- 以枸杞为试材，根据已知物种WRKY转录因子保守片段设计引物，采用RT-PCR方法，克隆枸杞WRKY基因片段，并对所得片段进行序列分析，同时利用Real-time PCR鉴定该基因在枸杞各器官的表达，以了解WRKY转录因子在枸杞中的功能。结果表明：从枸杞果实中获得了467 bp的cDNA片段，命名为LbWRKY (GenBank：KF181204)。序列分析表明，该序列含有WRKYGQK保守结构域，与番茄WRKY序列相似性是95%，与烟草wizz相似性是91%，表明已经成功克隆了枸杞WRKY基因片段。Real-time PCR表达分析显示，LbWRKY基因在枸杞果实中表达量最高，在叶中表达水平较低，在根中未检测到其表达。
- Abstract:
- Taking Lycium barbarum L.as material，primers were designed according to WRKY conserved transcription factor fragment form known species.Lycium barbarum L.WRKY gene fragments were cloned by RT-PCR method.Sequence of the fragments obtained was analyzed.The expression of Lycium barbarum L.in various organs was identified by Real-time PCR.They all contributed to research the function of WRKY transcription factor of Lycium barbarum L.The results showed that the cDNA fragment was 467 bp and named LbWRKY (GenBank：KF181204).Sequential analysis indicated that the sequence contained WRKYGQK conservative domain，and the similarity with Solanum lycopersicum WRKY and Nicotiana tabacum wizz were 95% and 91%，respectively.The results showed that WRKY gene sequence in Lycium barbarum L.was cloned successfully.Real time PCR analysis showed that the LbWRKY gene was highly expressed in fruit.The expression level reached the lowest level in leaf.LbWRKY didn’t express in root.
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备注/Memo
第一作者简介：王丽娟（1974-），女，博士，副教授，现主要从事植物基因工程研究工作。E-mail:Lijuanwang279@hotmail.com.