硫霉素生物合成基因簇的异源表达及其改造的开题报告.docx

硫霉素生物合成基因簇的异源表达及其改造的开题报告 摘要 硫霉素是一种重要的广谱抗生素，具有广泛的应用前景。硫霉素合成的生物合成基因簇（sul）中包括了多个基因，这些基因共同协同合成硫霉素。为了研究硫霉素的生物合成过程以及提高其生产效率，本研究计划异源表达硫霉素基因簇，并利用基因编辑技术进行改造，以达到提高硫霉素产量的目的。 本研究将构建一个自主复制、高效表达硫霉素基因簇的表达载体，并将其转化到大肠杆菌中。通过表达菌株的筛选和分析，确定最适合异源表达硫霉素基因簇的宿主菌株。为了提高硫霉素产量，我们将利用CRISPR-Cas9系统对sul基因簇进行基因编辑，找到主要限制硫霉素合成的关键因素，进行改造。最后，通过对表达菌株的鉴定和硫霉素的产量测定，验证我们的工作的有效性和可行性。 该研究的结果将为深入探究硫霉素合成机制，为硫霉素产业的发展提供重要的理论和实验依据。 关键词：硫霉素、基因编辑、异源表达、生物合成基因簇 Abstract Streptomycin is an important broad-spectrum antibiotic with broad application prospects. The biosynthetic gene cluster (sul) of streptomycin includes multiple genes that work together to synthesize streptomycin. In order to study the biosynthesis process of streptomycin and improve its production efficiency, this study plans to heterologously express the streptomycin gene cluster and use gene editing technology to modify it to achieve the goal of improving streptomycin production. This study will construct an expression vector that can autonomously replicate and efficiently express the streptomycin gene cluster, and transform it into E. coli. The most suitable host strain for heterologous expression of the streptomycin gene cluster will be determined through the screening and analysis of the expression strains. To improve the streptomycin production, we will use the CRISPR-Cas9 system to edit the sul gene cluster and find the key factors that limit the synthesis of streptomycin for modification. Finally, the effectiveness and feasibility of our work will be verified through identification of the expression strains and measurement of streptomycin production. The results of this study will provide important theoretical and experimental basis for in-depth exploration of the streptomycin biosynthesis mechanism and the development of the streptomycin industry. Keywords: streptomycin, gene editing, heterologous expression, biosynthetic gene cluster