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当前位置: 首页 > 文章 > 红肉苹果(Malus pumila var.niedzwetzkyana)MpMYB30基因的克隆及表达载体的构建 果树学报 2010,27 (4) 483-489
Position: Home > Articles > Cloning of MpMYB30 from Malus pumila var. niedzwetzkyana and construction of plant express vector Journal of Fruit Science 2010,27 (4) 483-489

红肉苹果(Malus pumila var.niedzwetzkyana)MpMYB30基因的克隆及表达载体的构建

作  者：
展蔷;张计育;佟兆国;董畅;章镇;渠慎春
单  位：
南京农业大学园艺学院
关键词：
红肉苹果;MpMYB30;生物信息学;表达载体构建
摘  要：
MYB转录因子是植物转录因子中最大的家族之一,在转录调节中起着多方面的重要作用。实验利用RACE技术从野生红肉苹果(Malus pumila var.niedzwetzkyana Schneid.)中获得了MpMYB30的全长序列,利用生物信息学的方法对其蛋白进行了分析和预测并将其连接到pCAMBIA-S1300+植物表达载体中。结果表明,该基因全长为1661bp,推导的氨基酸序列经系统发育分析与葡萄MYB30及蓖麻MYB基因相似性最高,命名为MpMYB30,其N端含有2个约55个氨基酸组成的MYB特征结构域。该基因推导的MpMYB30蛋白的分子式为C1790H2794N538O584S12,相对分子质量为41579.8ku,等电点为6.30,不存在信号肽和跨膜区域,蛋白质结构以α螺旋为主。利用XbalⅠ和SacⅠ对重组质粒进行酶切,并将其连接到pCAMBIA-S1300+载体上,通过抗性筛选和PCR酶切检测,证实了MpMYB30基因已经构建到植物表达载体pCAMBIA-S1300+上。
译  名：
Cloning of MpMYB30 from Malus pumila var. niedzwetzkyana and construction of plant express vector
作  者：
ZHAN Qiang,ZHANG Ji-yu,TONG Zhao-guo,DONG Chang,ZHANG Zhen,QU Shen-chun* (College of Horticulture, Nanjing Agriculture University, Nanjing 210095 China)
关键词：
Malus pumila var. niedzwetzkyana Schneid.; MpMYB; Bioinformatics; Expression vector
摘  要：
MYB, which plays an important role in many aspects, is one of the biggest families of transcription factor. Niedzwetzkyana schneid, a wild red flesh apple, was employed as the experimental material to clone MYB transcription factors. One of the MYB transcription factors, named MpMYB30 was cloned by RACE technology from the leaves of niedzwetzkyana schneid. The result showed that the total length of MpMYB30 gene was 1 661 bp. Bioinformatics analysis showed that the protein of MpMYB30 gene encoded had two MYB HTH DNA-binding domains, which shared compatibility with Vitis viniferna and Ricinus communis; the protein's molecular formula was C1790H2794N538O584S12 and the relative molecular weight was 41 579.8 ku;the isoelectric point was 6.30, and contained more α-helix in its secondary structure, but did not existed a signal peptide and transmembrane regions in its multi-peptide. The sequence of MpMYB30 gene was digested by XbalⅠand SacⅠ, then linked into pCAMBIA-S1300+vector, after that it was confirmed by resistance selected and PCR analysis.

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