川牛膝CoObgC基因克

发布时间：2018-04-03 19:07

  本文选题：川牛膝　切入点：ObgC　出处：《中国中药杂志》2016年14期

【摘要】：根据川牛膝基因组数据提供的基因序列合成特异性引物,利用常规PCR方法和cDNA末端快速扩增(RACE)技术克隆川牛膝Obg C(Gene Bank登录号KU847910)全长cDNA序列,克隆获得2 226 bp全长CoObgC序列,开放阅读框为1 818 bp,编码605个氨基酸序列,预测CoObgC蛋白相对分子质量为66.39 k Da,等电点p I 5.35,为稳定蛋白,并进行多重序列比对和构建系统进化树分析。以川牛膝actin为内参,采用实时荧光定量PCR(qRT-PCR)分析CoObgC基因在川牛膝根、茎、叶、花4种组织中表达特征,结果显示在叶片中表达丰度最高,其次为根、花、茎;构建p CABIA2300-CoObgC重组载体,利用农杆菌介导法在烟草中进行瞬时表达,激光扫描共聚焦显微镜观察显示川牛膝CoObgC定位于叶绿体。该研究为进一步解析Obg基因的结构和功能,开展川牛膝分子生物学研究奠定基础。
[Abstract]:Specific primers were synthesized from Genomic data of Achyranthes bidentata. The full-length cDNA sequence of Obg C(Gene Bank (KU847910) was cloned by routine PCR method and rapid amplification of cDNA terminal cDNA. The full-length CoObgC sequence was obtained.The open reading frame is 1 818 BP, encoding 605 amino acid sequences. The predicted molecular weight of CoObgC protein is 66.39kDa. the isoelectric point I 5.35 is a stable protein, and the multiple sequence alignment and phylogenetic tree analysis are carried out.The expression characteristics of CoObgC gene in root, stem, leaf and flower of Achyranthes bidentata were analyzed by real-time fluorescence quantitative analysis with actin as internal reference. The results showed that the expression of CoObgC gene was the highest in the leaves, followed by the roots, flowers and stems, and the recombinant vector of p CABIA2300-CoObgC was constructed.Transient expression was carried out in tobacco by Agrobacterium tumefaciens. Laser scanning confocal microscopy showed that CoObgC was located in chloroplast of Achyranthes bidentata.This study laid a foundation for the further analysis of the structure and function of Obg gene and the development of molecular biology of Achyranthes bidentata.
【作者单位】： 四川农业大学农学院;四川农业大学新农村发展研究院;
【基金】：国家“十二五”科技支撑计划项目(2011BAI13B02-7)
【分类号】：S567.239
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本文编号：1706517