白花丹参丙酮酸脱羧酶基因的克隆和表达分析_史仁玖.pdfVIP

• 90 • 中草药 Chinese Traditional and Herbal Drugs 第 44 卷 第 1 期 2013 年 1 月 白花丹参丙酮酸脱羧酶基因的克隆和表达分析 史仁玖，常正尧，王健美，王德才* 泰山医学院 医药生物技术研究所，山东 泰安 271000 摘 要：目的 获得白花丹参丙酮酸脱羧酶（SmPDC ）全长基因，分析该基因在白花丹参不同组织部位，以及缺氧胁迫处 理后的该基因表达差异。方法 利用 cDNA 文库筛选获得 SmPDC 基因全长，利用半定量 RT-PCR ，分析 SmPDC 基因在白 花丹参不同部位的表达情况，及缺氧处理条件下的表达情况。结果 获得的 SmPDC 基因由 2 190 个核苷酸组成，编码 605 4 个氨基酸，蛋白相对分子质量约 6.485 ×10 ，等电点pI 5.49 ；半定量RT-PCR 检测，该基因在丹参的根中表达量最高，其次 是茎和叶；缺氧胁迫处理会诱导该基因的表达，随胁迫时间延长表达量逐渐增加。结论 白花丹参 SmPDC 基因是 PDC 家 族新成员，其功能与植物耐缺氧代谢途径有关。 关键词：白花丹参；丙酮酸脱羧酶基因；克隆；RT-PCR ；缺氧胁迫 中图分类号：R282.12 文献标志码：A 文章编号：0253 - 2670(2013)01 - 0090 - 06 DOI: 10.7501/j.issn.0253-2670.2013.01.017 Cloning and expression analysis of pyruvate decarboxylase gene in Salvia miltiorrhiza SHI Ren-jiu, CHANG Zheng-yao, WANG Jian-mei, WANG De-cai Institute of Medicinal Biotechnology, Taishan Medical University, Tai’an 271000, China Abstract: Objective To obtain the fulllength of Salvia miltiorrhiza pyruvate decarboxylase (SmPDC) gene, to analyze the expression differences in various tissues of S. miltiorrhiza after anaerobic stress treatment. Methods The fulllength of SmPDC gene was isolated through sequencing cDNA library, and semi-quantitative RT-PCR was used to detect the gene expression levels. Results The fulllength of SmPDC cDNA has an open reading frame of 2 190 bp. The deduced amino acid sequence of SmPDC has 605 amino acid residues which form a 6.485 × 104 polypeptide with a calculated pI of 5.49. Semi-quantitative RT-PCR indicated that SmPDC gene was expressed at a high level in root, followed by stem and leaf of S. miltiorrhiza. Anaerobic stress could induce the expression of SmPDC gene and the expression was increased with the stress time elongating. Conclusion SmPDC is a new member of the PDC family and plays an important role in a