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确定人工合成的人肽抗生素hPAB β的抗菌活性。利用已知肽抗生素hBD 2的晶体结构坐标进行hPAB β的同源模建 ;设计hPAB β突变体 ;用PCR法生成hPAB β突变体基因 ,分别构建转移载体和重组杆状病毒。结果表明 ,合成肽hPAB β在正确复性后具有较好的杀菌活性 ,将 4种突变体基因插入转移载体pFASTHTa ,筛选阳性重组子pFAST hPAB β并转化DH10Bac大肠杆菌 ,获得了重组杆状病毒 ,为筛选活性强而结构更为简单的hPAB β突变体奠定了基础
The antibacterial activity of the synthetic human antibiotic hPAB β was determined. Homology modeling of hPAB β was performed using the crystal structure coordinates of the known peptide antibiotic hBD 2. The hPAB β mutant was designed. The hPAB β mutant gene was generated by PCR to construct the transfer vector and the recombinant baculovirus. The results showed that the synthetic peptide hPAB β had good bactericidal activity after correct refolding. Four kinds of mutant genes were inserted into the transfer vector pFASTHTa, the positive recombinant pFAST hPAB β was screened and transformed into DH10Bac E. coli, and the recombinant baculovirus was obtained. Which laid the foundation for the screening of hPABβ mutant with stronger activity and simpler structure