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从蜡梅中克隆α-半乳糖苷酶基因,通过对其酶学性质及基因表达分析,为研究蜡梅的低温适应的分子机理提供参考.通过PCR扩增获得α-半乳糖苷酶基金(CpGAL),构建了该基因的原核表达载体pET28a-CpGAL并转化到大肠杆菌BL21(DE3)中,诱导表达并分离纯化得到融合蛋白.利用SYBR GreenⅠ实时荧光定量PCR检测CpGAL基因在腊梅不同发育时期的表达差异.
The α-galactosidase gene was cloned from Chimonanthus praecox, with reference to its enzymatic properties and gene expression analysis for studying the molecular mechanism of Chimonanthus praecox in cold adaptation.A-Galactosidase Fund CpGAL) was constructed and the prokaryotic expression vector pET28a-CpGAL of the gene was constructed and transformed into E. coli BL21 (DE3) to induce expression and separation and purification of the fusion protein.Using SYBR Green Ⅰ real-time quantitative PCR to detect CpGAL gene expression in different developmental stages Differences in expression.