地黄类RghBNG基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of Rgh BNG-like Gene in Rehmannia glutinosa

Rgh BNG基因是一种地黄(Rehmannia glutinosa)响应非生物胁迫和植物生长调节剂的基因。类Rgh BNG基因是一个与Rgh BNG基因核酸序列同源的地黄基因。克隆了地黄类Rgh BNG基因,预测其编码蛋白质的性质、结构和功能。用RNA提取试剂盒提取地黄总RNA,反转录成c DNA,根据已知地黄的Rgh BNG基因碱基序列设计上、下游引物,以c DNA为模板,利用PCR扩增产生包括Rgh BNG基因和类Rgh BNG基因在内的5个c DNA片段,其中类Rgh BNG基因全长1 974 bp,包含一个486 bp的ORF,编码161个氨基酸残基组成的蛋白质;蛋白质等电点为9.45,分子量为18.6 k Da,二级结构中延伸链占34.78%,无规则卷曲占33.54%,α-螺旋占21.74%,β-螺旋占9.94%,有两个可能的跨膜区,参与翻译和脂肪酸代谢。

Rgh BNG gene is a novel abiotic stress and plant growth regulator-responsive gene from Rehmannia glutinosa,while Rgh BNGLgene is Rgh BNG-like gene homologous to Rgh BNG gene at the nucleotide level. Rgh BNG-like gene was successfully cloned from Rehmannia glutinosa,and predicted byabioinformatics softwares. The total RNA was extracted from Rehmannia glutinosa leaves with RNA extraction kit,and then used as reverse transcriptional template to clone Rgh BNG-like gene with the same primer pair as that for Rgh BNG gene. Rgh BNG-like gene was 1 974 bp long,including a 486 bp ORF,encoding for161 amino acid residues. The protein isoelectric point and molecular weight are 9.45 and 18.6 k Da. Its secondary structure consists of 34.78% extended strand,33.54% random coil,21.74% alpha helix and 9.94% β-turn. It has two transmembrane helical segments and takes part in translation and fatty acid metabolism.