【摘要】 为构建HMG-17原核表达系统,研究HMG-17抗菌功能与结构的关系,提取人LAK细胞总RNA,设计合成相应引物,应用RT-PCR扩增HMG-17和HMG-17α螺旋结构成熟肽cDNA,以pGEX-1λT为载体,构建HMG-17和HMG-17α螺旋结构基因大肠杆菌表达载体。该载体表达的融合蛋白GST-HMG-17和GST-HMG-17α经亲合层析、凝血酶酶切、AU-PAGE胶回收后获得高度纯化的HMG-17和HMG-17α肽,对纯化产物进行MEC、MIC、MBC等试验,与杀菌活性较强的抗菌肽HNP-1、NP-1进行活性对比,证明其具有较强的杀菌活性,且具有盐敏感性。并证明了HMG-17的α结构域是HMG-17的主要抗菌功能区域。更多还原

【Abstract】 Total RNA was extracted from human LAK cell, and a cDNA encoding mature peptide HMG-17 and its α helix domain was amplified by RT-PCR. The recombinant prokaryotic expression vector pGEX-1λT-HMG-17 and pGEX-1λT HMG-17α helix was constructed. Using affinity chromatography, thrombin cleaving and AU-PAGE elution,we obtained the purified HMG-17. Analyses of MIC, MEC and MBC indicated that HMG-17 and HMG-17α had strong antibacterial activity. MIC of the α-helic domain was almost the same as that of HMG17, suggesting that the α-helic structure would be essential for the antibacterial activity of HMG-17.更多还原