中华补血草Syntaxin基因的克隆

目的]对中华补血草Syntaxin基因进行克隆。方法]以中华补血草叶片为材料,提取总RNA并进行反转录反应。依据已知Syntaxin基因5'端EST序列信息设计巢式引物,以反转录得到的cDNA为模板,利用2轮PCR扩增cDNA3'末端(3'RACE),获得Syntaxin基因3'端序列。结果]获得1090bp的DNA片段。分析表明,该片段编码LsSyntaxin全长基因,其中开放阅读框长816bp,编码271个氨基酸。该基因编码的Syntaxin蛋白相对分子量为30254.3Da,理论等电点为5.55。结论]为研究Syntaxin基因在补血草中的功能及其在补血草泌盐过程的作用奠定了基础。

Cloning of Syntaxin Gene in Limonium sinense Kuntze

Objective] The aim was to clone Syntaxin genes in Limonium sinense Kuntze. Method] Limonium sinense Kuntze leaves were used as materials and total RNA was extracted and transcribed reversely. Nested primers were designed as per EST sequences at gene 5’ region of Syntaxin, and cDNA obtained through reverse reaction was taken as the template. Sequences of Syntaxin gene at 3’ region were achieved through two rounds of PCR amplifications. Result] DNA fragments (1 096 bp) were obtained. For LsSyntaxin, open reading frame (ORF) was 816 bp and the encoded amino acids were 271. The relative molecular weight of Syntaxin was 30 254.3 Da and isoelectric point in theory was 5.55. Conclusion] Syntaxin genes from Limonium sinense Kuntze were cloned. The research laid foundation for study of Syntaxin gene function in Limonium sinense Kuntze and salt-secreted process.