摘要
为探明萼脊兰EF1α基因在生长发育过程中的生理功能和作为内参基因的稳定性及适用性,根据NCBI中植物翻译延长因子同源核苷酸保守序列设计简并引物,以萼脊兰叶片为试验材料,采用RNAprep多糖多酚植物总RNA提取试剂盒提取RNA,利用RT-PCR技术克隆翻译延伸因子EF1α,并进行生物信息学分析。结果表明:EF1α基因长620bp,编码206个氨基酸,编码的蛋白为亲水性蛋白,相对分子质量为23.16KD,等电点为8.50;经二级结构分析,α-螺旋占29.61%,延伸链占31.55%,无规则卷曲占38.83%;亚细胞定位在细胞质;EF1α基因与朵丽蝶兰杂交品种翻译延伸因子蛋白的一致性较高,进化距离最近。在GenBank登录号为KT223116。
To explore gene EF1αin the process of growth and development,physiological functions and lay a foundation as internal genetic stability and applicability,EF1αof S.japonica was cloned and sequence analyzed.According to the NCBI login plant translation elongation factor homologous conserved sequences of nucleotide primers to design degenerate primers,S.japonica leaves as experimental materials,RNAprep pure plant kit was used to extract RNA,and use RT-PCR technology to clone the translation elongation factor EF1 a.Through a series of bioinformatics analysis,the results showed that the gene sequence was 620 bp,encoding 206 amino acid residues,was a hydrophilic protein,molecular weight was 23.16 KD,isoelectric point was 8.50.Secondary structure analysis showed that the alpha helix occupied 29.61%,extended strand accounted for 31.55%,Random coil accounted for 38.83%.Subcellular located in cytoplasm.Protein sequence and phylogenetic analysis showed that it had high consistency and short evolutionary distance with the Doritaenopsis hybrids of translation elongation factor protein.The authors registered them into GenBank（respective accession number：KT223116）.
出处
《贵州农业科学》
CAS
2016年第5期5-8,共4页
Guizhou Agricultural Sciences
基金
河南省科技攻关项目"蝴蝶兰的分子育种"(092102110128)
郑州市普通科技攻关项目"蝴蝶兰新品种选育研究"(141PPTG G420)
关键词
萼脊兰
EF1α
基因克隆
二级结构
Sedirea japonica
EF1α
gene clone
secondary structure
