CAJ | 학술논문

基于GenBank报道的近缘物种C4H和CAD基因cDNA序列，应用Primer Premier 5.0软件设计2对PCR扩增引物。用CTAB-LiCl法提取芒草总RNA，经反转录后合成cDNA，应用RT-PCR方法成功扩增出MsC4H和MsCAD基因片段并克隆到pMD18-T载体。测序结果表明MsC4H和MsCAD基因片段长度分别为305 bp和269 bp，编码101个和51个氨基酸。Blast分析结果显示克隆序列与其它植物的C4H及CAD基因具高同源性。已将克隆的MsC4H和MsCAD基因cDNA序列提交至GenBank数据库，其注册号分别为JQ598686和JQ598683。
기우GenBank보도적근연물충C4H화CAD기인cDNA서렬，응용Primer Premier 5.0연건설계2대PCR확증인물。용CTAB-LiCl법제취망초총RNA，경반전록후합성cDNA，응용RT-PCR방법성공확증출MsC4H화MsCAD기인편단병극륭도pMD18-T재체。측서결과표명MsC4H화MsCAD기인편단장도분별위305 bp화269 bp，편마101개화51개안기산。Blast분석결과현시극륭서렬여기타식물적C4H급CAD기인구고동원성。이장극륭적MsC4H화MsCAD기인cDNA서렬제교지GenBank수거고，기주책호분별위JQ598686화JQ598683。
Based on cDNA sequences encoding cinnamate-4-hydroxylase (C4H) and Cinnamoyl alcohol dehyd-rogenase (CAD) isolated from several monocots reported in GenBank, two pairs of PCR primers were designed with Primer Premier 5.0 software. The total RNA was extracted from Miscanthus sinensis according to CTAB-LiCl method and then cDNA was synthesized by reverse transcription. PCR products, which were named as MsC4H and MsCAD, were successfully obtained by reverse transcription polymerase chain reaction (RT-PCR) and then cloned into vector pMD18-T. The positive clones identified by plasmid PCR were sequenced. The sequencing results revealed that MsC4H and MsCAD contain 305 and 269 base pairs, which encode 101 and 51 amino acids, respectively. Blast analysis showed that the cloned nucleotide sequences and their corresponding amino acids are highly homologous to many C4H and CAD genes and their encoding proteins from different plant species. The sequences of MsC4H and MsCAD had been submitted to GenBank database, whose accession numbers are JQ598686 and JQ598683, respectively.