人参HMGR基因的克隆与序列分析

以人参根RNA为模板,根据其他植物HMGR基因序列保守区设计特异性简并引物,进行RT-PCR扩增。纯化回收HMGR基因RT-PCR产物与pMD18-T载体连接,再转化到JM109大肠杆菌中进行克隆,对阳性克隆菌重组质粒进行测序和序列分析。结果表明:人参HMGR核心片段的大小为458bp,与其他植物核苷酸序列有较高的同源性,依次为杜仲86.1%、南京椴83.8%、长春花83.2%、马铃薯82.5%、苹果82.1%、景烈白兰81.6%、肾叶橐吾81.6%、南苍术81.0%、红豆杉80.4%、水稻80.3%。

Cloning and Sequence Analysis of HMGR Gene in Ginseng

Using RNA from ginseng roots as templates, designing specific degenerate primers based on conserved HMGR gene from various plants, RT－PCR was performed. The products of RT－PCR were recovered, purified and cloned into pMD18 T vector, then transformed into competent Escherichia coli JM109. Inserts  included in the plasmid were sequenced and analyzed. The results showed that the gene of HMGR was 458 bp, the nucleotide sequence showed high homology with other plants. The identities of nucleotide sequences were Cortex Eucommiae 86.1％,Tilia miqueliana Maxim 83.8％,Catharanthus roseus (L.) Don 83.2％, Solanum tuberosum82.5％, Malus pumila Mill　82.1％,Michelia chapensls 81.6％, Ligularia hodgsoni　Hook. var. sutchuenensis (Franch.) Henry　81.6％, Rhizoma Atractyl ods 81.0％, Taxaceae　80.4％,Oryza sativa 80.3％.