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应用新的融合方法制备抗菌肽CMIV变体;了解重组型抗菌肽变体对几株临床耐药菌的抗性。通过基因工程的手段,将合成的抗菌肽CMIV的cDNA与根据金黄色葡萄球菌A蛋白(SPA)的IgG结合结构域设计并合成的ZZ结构域基因融合,在大肠杆菌细胞中表达;用制备的CMIV变体进行抑菌圈试验和液相试验。结果表明5ul(10umol/L)抗菌肽可在大肠杆菌K12D31平板上产生直径15mm的抑菌圈;抑制50%大肠杆菌生长的抗菌肽浓度约0.2umol/L。6种青霉素抗性的革蓝氏阴性菌的抑菌圈直径在6~13mm之间;而同样的条件下,对革蓝氏阳性菌——金黄色葡萄球菌的作用很弱。用新的融合方法可通过大肠杆菌体系制备有活性的抗菌肽变体,重组型抗菌肽有可能成为新一代的抗菌素
The new fusion method was used to prepare the antimicrobial peptide CMIV variant. The resistance of the recombinant antimicrobial peptide variant to several clinical drug-resistant strains was also studied. The gene of synthetic antibacterial peptide CMIV was genetically engineered and fused with the ZZ domain designed and synthesized according to the IgG binding domain of Staphylococcus aureus Protein A (SPA), and expressed in E. coli cells. CMIV variants were tested for inhibition zone and liquid phase. The results showed that 5ul (10umol / L) antibacterial peptide can produce antibacterial ring with a diameter of 15mm on E. coli K12D31 plate. The concentration of antibacterial peptide inhibiting growth of 50% Escherichia coli was about 0.2umol / L. The antibacterial circle diameter of 6 kinds of penicillin-resistant gram-negative bacteria was between 6 ~ 13mm; while under the same conditions, the effect on Gram-positive bacteria Staphylococcus aureus was very weak. With the new fusion method, active antibacterial peptide variants can be prepared by the E. coli system. Recombinant antibacterial peptides may become a new generation of antibiotics