报道了鲑鱼降钙素基因植物表达载体的构建及其向烟草的遗传转化。由鲑鱼降钙素的链霉菌表达载体 p MS6 80 ,通过限制性内切酶 Bam H 和 H ind 双酶切后将其克隆至中间载体 p ART7,获得中间表达载体p ART7- s CT,从而给目的基因加上启动子和终止子 ,将限制性内切酶 N ot1酶切 p ART7- s CT得到的带有启动子和终止子的目的基因片段克隆至载体 p ART2 7,得到鲑鱼降钙素基因的植物表达载体 p ART2 7- s CT。通过农杆菌介导的叶盘法将其转至烟草 ,经 PCR检测已得到转基因植株。
[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 based on EST sequences at 5’ region of Syntaxin, and cDNA obtained through reverse reaction was taken as the template. Sequences of Syntaxin gene at 3’ region were obtained 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 the study on Syntaxin gene function in Limonium sinense Kuntze and salt-secreted process.
