枣树内参基因ZjH3的克隆与筛选

Cloning and Selection of Housekeeping Gene ZjH3 for Ziziphus jujuba

为了克隆和筛选适合于枣树(Ziziphus jujuba Mill.)基因表达研究用的内参基因,利用枣结果枝cDNA文库ESTs序列,克隆获得4条其它物种常用内参基因的同源基因:组蛋白H3的同源基因全长序列,命名为ZjH3(Ziziphus jujubaH3,GenBank登录号:EU916201);肌动蛋白的同源基因片段,命名为ZjAT1(Ziziphus jujubaACT,GenBank登录号:EU251882);泛素延伸蛋白同源基因全长序列,命名为ZjUBQ(Ziziphus jujubaUBQ,GenBank登录号:EU916200);翻译延伸因子的同源基因片段,命名为ZjEF1(Ziziphus jujubaEF,GenBank登录号:EU916202)。在相同的条件下,用半定量RT-PCR方法分析了这4个潜在内参基因在枣不同生长时期的结果枝及其茎尖和不同器官中的表达,结果表明,ZjAT1、ZjUBQ和ZjEF1基因的表达不恒定,不适合用作枣树内参基因;而ZjH3基因在不同发育阶段的结果枝及其茎尖、根、茎、叶、芽、花蕾、花、幼果、膨大果实和种子中均表达稳定,适合于用作枣树的内参基因。

To select reliable housekeeping gene used in gene expression studies of chinese jujubes,four potential housekeeping genes were isolated from cDNA Library in the Fruit-Bearing Shoot of Ziziphus jujuba Mill.including a homologue gene of histone3 designated ZjH3（GenBank accession number：EU916201）,a homologue gene of β-actin designated ZjAT1（GenBank accession number：EU251882）,a homologue gene of ubiquitin extension protein designated ZjUBQ（GenBank accession number：EU916200）,and a homologue gene of translation elongation factor 1α designated ZjEF1（GenBank accession number：EU916202）.Expression of these genes in the fruit-bearing shoot during different growth period and its apices tissue,vegetative shoot,and different other tissues were analyzed by using semi-quantitative RT-PCR under the same conditions.The results showed that ZjAT1,ZjUBQ and ZjEF1 gene revealed unstable expression and were not suitable as housekeeping genes,but ZjH3 was the most suitable housekeeping gene for Ziziphus jujuba,based on its stable expression in different organs.