玉米光周期敏感基因ZmELF4的克隆及其亚细胞定位

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。

Gene Cloning and Subcellular Location of Photoperiod Sensitive Gene ZmELF4 in Maize

Primers were designed according to conserve sequences of ELF4 of arabidopsis thaliana.Using RT-PCR, the full-length cDNAs of the homologous gene of ELF4 in maize was obtained from a photosensitive tropical maize inbred line CML288(GenBank accession number was HQ009862). Sequence analysis indicated that the full-length cDNAs was composed of 683 bp sequences including an open reading frame (ORF) of 432 bp region which encoded 144 amino acids. The ZmELF4 gene shared a amino acid sequence homology of 62% with Arabidopsis thaliana, and 75% with rice. In addition, they all had a highly conserved domain of unknown function, named DUF1313. Using recombinant DNA technology, the plant over experssion vector and fusion expression vector were constructed, which respectively named as pBI-ZmELF4 and pGIT-ZmELF4-GFP, these laid the foundation for further gene function validation by transgenic technology. We had used fusion expression vector to carry out transient expression experiments in onion epidermal, the results of this experiment showed that ZmELF4 was located in the nucleus, this explained the role of the gene was in the nucleus.