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在分析和筛选我们前期建立的地黄转录组文库、抑制消减杂交文库及地黄相关EST文库基础上,利用拼接、延伸及PCR方法,成功克隆到一个全长2681bp的Ca2+-ATPase基因,含有2007bp完整的开放阅读框,编码568个氨基酸。分析表明,地黄Ca2+-ATPase属于内质网型ATPase,与大豆和葡萄有较近亲缘关系,Ca2+-ATPase含有Ca2+离子转运区、ATP结合区等保守区。随着生长,正茬地黄Ca2+-ATPase表达量逐渐升高,而连作地黄中的Ca2+-ATPase基因在块根伸长期表达突然升高,之后降低。
Based on the analysis and screening of the cDNA library of Rehmannia glutinosae, suppression subtractive hybridization library and Rehmannia glutinosa EST library, we successfully cloned a 2681bp full-length Ca2 + -ATPase gene with complete 2007bp Open reading frame, encoding 568 amino acids. Analysis showed that Rehmannia glutinosa Ca2 + -ATPase belongs to the endoplasmic reticulum ATPase, and soybean and grape have a closer genetic relationship, Ca2 + -ATPase contains Ca2 + ion transport region, ATP binding region and other conserved regions. With the growth, Ca up the Rehmannia glutinosa Ca2 + -ATPase expression gradually increased, and even the Rehmanniae Ca2 + -ATPase gene expression in the tuberous elongation of a sudden increase, then decreased.