Affiliation:
1. State Key Laboratory of Biobased Material and Green Papermaking, Shandong Academy of Sciences, Qilu University of Technology, Jinan 250353, China
2. Shandong Provincial Key Laboratory of Microbial Engineering, School of Bioengineering, Shandong Academy of Sciences, Qilu University of Technology, Jinan 250353, China
Abstract
Plasmids are commonly used tools in microbiology and molecular biology and have important and wide-ranging applications in the study of gene function, protein expression, and compound synthesis. The complex relationship between necessary antibiotic addition, compatibility between multiple plasmids, and the growth burden of host bacteria has plagued the wider use of compatibility plasmids. In this study, we constructed the terminal polymerization pathway of PSA by exogenously expressing the neuA, neuD, and neuS genes after the knockdown of Eschesrichia coli BL21 (DE3). Duet series vectors were utilized to regulate the expression level of neuA, neuD, and neuS genes to study the gene expression level, plasmid copy number growth burden, pressure of antibiotic addition, stability of compatible plasmids, and the level of expression stability of exogenous genes, as well as the effect on the biological reaction process. The results showed that the three genes, neuA, neuD, and neuS, were enhanced in the recombinant strain E. coli NA-05, with low copy, medium copy, and high copy, respectively. The effect of PSA synthesis under standard antibiotic pressure was remarkable. The results of this thesis suggest the use of a Duet series of compatible expression vectors to achieve the stable existence and co-expression of multiple genes in recombinant bacteria, which is a good reason for further research.
Funder
Pilot Project for the Integration of Science, Education and Industry, Qilu University of Technology, Shandong Academy of Sciences
Shandong Province Science and Technology SMES Innovation Ability Improvement Project
Dezhou Major Science and Technology Innovation Project and Rizhao Key Research and Development Plan
Subject
Plant Science,Biochemistry, Genetics and Molecular Biology (miscellaneous),Food Science
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