GNA成熟蛋白基因亚克隆及其原核表达载体构建

Subcloning of GNA mature protein (MGNA) gene and its construction of expression vector in E.coli

雪花莲外源凝集素 (GNA)对刺吸式昆虫和某些咀嚼式昆虫以及多种线虫均有毒性。从含GNA前体蛋白基因的质粒中亚克隆出GNA的成熟蛋白基因MGNA ,将MGNA基因插入大肠杆菌表达载体pET2 2b的不同位点 ,再经测序验证 ,得到了三种不同表达形式的GNA原核表达载体 :2 2bG1 (分泌型融合GNA蛋白 )、2 2bG2 (包涵体型GNA蛋白 )、2 2bG3(分泌型天然GNA蛋白 ) ,这为进一步在大肠杆菌中表达GNA和将GNA制成生物农药奠定了基础。

Snowdrop lectin mature protein is toxic to sap sucking insects and some chewing pests and many kinds of nematode.Mature protein gene of snowdrop lectin (MGNA) was subcloned in T vector from plasmid containing its precursor protein gene.MGNA was inserted downstream of T7 promoter at three different sites of E.coli expression vector pET22b(+),resulting in the construction of three prokaryotic expression vectors p22bG1(secretion of fusion GNA),p22bG2(Inclusion body) and p22bG3 (secretion of native GNA). The three vectors give different types of MGNA product expressed in E.coli. This research will be the base for MGNA expression in E.coli and make it into bio-pesticide.