CAJ | 학술논문

采用RACE方法从曼地亚红豆衫中成功克隆到Tm13AH基因.该基因包含一个1455 bp的开放阅读框,编码一个具有485个氨基酸,大小为54.6 kD,等电点为pH 8.81的蛋白质.序列同源性比对表明:它与其他种的羟化酶具有较高的同源性,为P450羟化酶家族的成员.组织特异性表达分析结果表明:Tm13AH在叶中表达量高,在茎中表达量低,在果实中无表达.Tm13AH基因的克隆和分析为进一步在分子水平上研究其在紫杉醇代谢途径中的功能奠定了基础.
채용RACE방법종만지아홍두삼중성공극륭도Tm13AH기인.해기인포함일개1455 bp적개방열독광,편마일개구유485개안기산,대소위54.6 kD,등전점위pH 8.81적단백질.서렬동원성비대표명:타여기타충적간화매구유교고적동원성,위P450간화매가족적성원.조직특이성표체분석결과표명:Tm13AH재협중표체량고,재경중표체량저,재과실중무표체.Tm13AH기인적극륭화분석위진일보재분자수평상연구기재자삼순대사도경중적공능전정료기출.
A full-length cDNA encoding Taxane 13α-hydroxylase (designated as Tm13AH), which catalyzes the second hydroxylation step in the Taxol biosynthetic pathway, was isolated from Taxus x media by rapid amplification of cDNA ends (RACE). The full-length cDNA of Tm13AH had a 1455 bp open reading frame (ORF) encoding a protein of 485 amino acid residues. The deduced protein had an isoelectric point (pI) of 8.81 and a calculated molecular weight of about 54.6 kD. Sequence comparison showed that Tm13AH had high identity to hydroxylases of other plant species reported previously. Transcription pattern analysis revealed that Tm13AH had high transcription in leaves, low in stems and no expression could be detected in fruits. The cloning and characterization of Tm13AH gene will enable us to further understand the role of Tm13AH involved in Taxol biosynthetic pathway in Taxus x media at molecular level.