白花丹参丙酮酸脱羧酶基因的克隆和表达分析

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个氨基酸,蛋白相对分子质量药6.485×104,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.

Cloning and expression analysis of pyruvate decarboxylase gene in Salvia miltiorrhiza

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 anaerobic respiration pathway.