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以热带玉米自交系CML288为供试材料,根据拟南芥ELF4基因保守序列设计引物,采用RT-PCR方法克隆了ELF4在玉米上同源基因的全长cDNA序列(GenBank上的登录号为HQ009862)。序列分析表明,该基因cDNA序列全长683 bp,开放阅读框为432 bp,编码了144个氨基酸,与拟南芥、水稻的氨基酸序列同源性分别高达62%和75%,此外它们还共同包含一个高度保守的DUF1313未知功能结构域。利用基因重组技术分别构建了能够高效表达的过量表达载体pBI-ZmELF4和带有GFP标记的融合表达载体pGIT-ZmELF4-GFP,利用融合表达载体进行洋葱表皮瞬时表达实验,结果将ZmELF4定位于细胞核内,说明该基因在细胞核内起作用。
Based on the conserved sequence of Arabidopsis thaliana ELF4 gene, the tropical maize inbred line CML288 was used as the test material, and the full-length cDNA sequence of the homologous gene of ELF4 in maize was cloned by RT-PCR (GenBank accession number: HQ009862 ). Sequence analysis showed that the cDNA sequence of the gene was 683 bp in length and 432 bp in open reading frame, encoding a protein of 144 amino acids. The deduced amino acid sequence shared 62% and 75% identity with Arabidopsis thaliana and rice, respectively. In addition, Contains a highly conserved DUF1313 unknown functional domain. The overexpression vector pBI-ZmELF4 and the GFP-tagged fusion expression vector pGIT-ZmELF4-GFP were constructed respectively by gene recombination technology. The transient expression of onion epidermis was confirmed by the fusion expression vector. The results showed that ZmELF4 was localized in the nucleus , Indicating that the gene plays a role in the nucleus.