甘蔗蔗糖磷酸合成酶磷酸化位点的突变及相应表达载体的构建

Site-directed mutagensis of the phosphorylation site of sucrose phosphate synthase from Saccharum officinarum and constructed the relation expression vector

用Clontech公司的定点突变试剂盒TransformerTMSite-Directed Mutagenesis Kit对克隆载体pMD18T-SPS3L上的SPS基因SPS3L进行定点突变,使其459位的丝氨酸密码子分别突变为编码苏氨酸、丙氨酸和谷氨酸的密码子。然后将突变后的和未突变的SPS3L基因分别克隆至表达载体质粒pCAMBIA1301中,构建重组质粒pCAMBIA1301-SPS3L。经测序鉴定,对SPS3L基因的定点突变成功,构建表达载体pCAMBIA1301-SPS3L成功。

The SPS3L gene of sucrose phosphate synthase（SPS） on the vector pMD18T-SPS3L was Site-directed Mutagensised by the TransformerTM Site-Directed Mutagenesis Kit（Clontech）,the Ser condon which at 456 to 459 was respectively mutagensised into three condons（Thr,Ala,Glu）.The mutagensised and unmutagensised SPS3L were cloned into expression vector pCAMBIA1301-SPS3L.According to the sequenced result,the Site-directed Mutagensis was correct and the expression vector pCAMBIA1301-sps3L was constructed successfully.