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《Acta Academiae Medicinae Militaris Tertiae》 2006-22
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Reconstruction of engineered bacteria in preparing recombinant human peptide antibiotic β

ZHANG Chun, CONG Yan-guang, HU Fu-quan, RAO Xian-cai (Department of Microbiology, College of Medicine, Third Military Medical University, Chongqing 400038, China)  
Objective To reconstruct the new engineered bacteria expressing hPAB-β triploids so as to improve the outputs of recombinant human peptide antibiotic β. Methods The recombinant plasmid pQE31-hPAB-β(3) was transformed into E. coli. M15 to screen the new engineered bacteria expressing hPAB-β triploids. The stabilities of phPAB-β(3)/M15 were observed in continuous cultures. The expression levels of the fusion peptides of interest and the bacterial yields of the new engineered bacteria phPAB-β(3)/M15 were compared with that of phPAB-β(3)/JM109 in different fermentation scales. Results Genetic stability of the recombinant plasmid and phPAB-β(3)/M15 was 100 after 10 passages. Take bacterial yields into account, the new engineered bacteria phPAB-β(3)/M15 was better than phPAB-β(3)/JM109 at the similar expression levels of the target proteins by “t” test analysis (P0.01). The yields of fermentation reached 55.15 g/L in 10 L fermenter, and the expression levels were up to 30.2 at 5th h after induction. During the fermentation, the recombinant plasmids were stable in culture. Conclusion The new engineered bacterium phPAB-β(3)/M15 successfully constructed was more consummate and effective on the basis of the original process.
【Fund】: 国家自然科学基金资助项目(30171119);; 全军医学科学技术研究“十五”计划基金重点课题(01Z070);; 国家“863”项目(2002AA214211)~~
【CateGory Index】: Q789
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