山药 DoARID4基因克隆与表达特异性分析

以''铁棍山药''为材料,根据同源克隆的方法获得''铁棍山药'' DoARID4序列,并对其氨基酸序列进行生物信息学分析,并构建系统进化树,通过qRT-PCR试验对该基因进行初步的组织特异性表达分析初步探究 DoARID4在''铁棍山药''生长发育以及扁茎发生中的调控作用,为ARID基因家族的进一步研究提供理论依据。结果表明,克隆得到一条长731 bp的条带,将测序结果进行Blast比对,发现其与芋头、莲藕ARID4序列相似性最高,因此将该基因命名为 DoARID4。 DoARID4可编码211个氨基酸,其蛋白质分子质量为  23.73 ku;其蛋白分子式为C₁₀₃₅H₁₆₀₆N₃₀₄O₃₀₅S₁₇,是一种脂溶性很强、非跨膜、亲水性蛋白;可能分布于细胞核及线粒体中,并且可能通过丝氨酸或苏氨酸磷酸化进行细胞信号的传导功能。 DoARID4在地上茎与地下茎的相对表达量较高,而在叶片中相对表达量较低,推测 DoARID4在山药茎和根的畸形发育中发挥重要作用。

Cloning and Expression Specific Analysis of DoARID4 Gene from Dioscorea oppositifolia L.

Using ''Tiegun yam'' as material, the DoARID4 sequence was obtained by the method of homologous cloning, and its amino acid sequence was analyzed by bioinformatics, a phylogenetic tree was constructed.by qRT-PCR test, preliminary tissue-specific expression analysis of the gene was conducted, and the regulatory role of DoARID4 in the growth and development of ''Tiegun yam'' and the development of flat stems were explored. This study provides a theoretical basis for further research on the ARID gene family. The results showed that a gene with 731 bp was cloned, and the sequencing results were compared with Blast. It was found that taro had the highest ARID4 sequence similarity with lotus root, so the gene was named DoARID4. DoARID4 encoded 211 amino acids with a protein molecular mass of 23.73 ku; its protein molecular formula was C₁₀₃₅H₁₆₀₆N₃₀₄O₃₀₅S₁₇, which was a highly fat-soluble, non-transmembrane, and hydrophilic protein; it may be distributed in the nucleus and mitochondria, and may pass through serine or threonine phosphorylation performs cell signal transduction function. The relative expression level of DoARID4 in above-ground-stems and underground-stems were relatively high, but the relative expression level was low in leaves. It is speculated that DoARID4 plays an important role in the abnormal development of yam stems and roots.