多花水仙HMGS基因的克隆

Cloning of HMGS gene from flowers of Narcissus tazetta

以多花水仙花瓣抑制消减杂交文库获得的羟甲基戊二酰辅酶A合酶(HMGS)基因片段为基础,采用cDNA末端快速扩增(RACE)技术从黄花水仙2号和金盏银台中各克隆一条HMGS基因,两基因均含有一个1413 bp的开放阅读框,编码470个氨基酸,但存在2个氨基酸差异.氨基酸序列分析表明:黄花水仙2号与葡萄、大豆、蓖麻和玉米HMGS基因的氨基酸相似系数分别为83%、82%、82%和81%.以水仙Actin基因为内参基因,采用荧光定量PCR方法分析HMGS基因在两品种的表达水平,结果表明:两品种HMGS基因的表达水平差异并不明显.

Two 3-hydroxy-3-methylglutaryl-CoA synthase （HMGS） genes were cloned from Narcissus tazetta var.Huanghuashuixian H and Jinzhanyintai by rapid amplification of cDNA ends （RACE）. The open reading frame （ORF） in both ofthe gene encompassed 1413 bp encoding a polypeptide of 470 amino acids, and there are two amino acids different between them.Sequencing analysis showed that the amino acid sequence of N. tazetta was 83%, 82%, 82%, 81% homologous with HMGS genesfrom Vitis vinifera, Glycine max, Ricinus communis, Zea may respectively. The relative expression levels of HMGS at two varietieswere analyzed by qRT-PCR and Actin was used as reference gene. The qRT-PCR results showed that HMGS relative expression lev-els at the two varieties are not obviously differential.