随着多耐药和极度耐药结核分枝杆菌增多，亟待找到新的结核病治疗药物。核糖体作为多种抗生素的作用靶点，对它的功能研究具有重要意义。致病性结核分枝杆菌生长慢、培养条件严苛，常以生长快、非致病的耻垢分枝杆菌进行替代研究，但两者核糖体数量和密度、代谢多样性存在显著差异。核糖体与代谢调控、生长速度之间存在怎样内在的联系？为探索此问题，我们采用了合成生物学的研究策略：通过构建来学习。现已将结核分枝杆菌散布在染色体24个位点的核糖体基因拼接成簇，拟将其整合进耻垢分枝杆菌后，再通过内源基因的系统性敲除实现耻垢分枝杆菌核糖体的替换。在此过程中，探究核糖体基因拷贝数与生长速度、基因染色体排布与表达调控、核糖体异质与翻译调控之间的关系。该研究可能筛选出一批功能差异的分枝杆菌核糖体基因，还可能获得大部分或全部核糖体基因被结核分枝杆菌同源物替换的新底盘，为将来针对分枝杆菌的药物筛选和核糖体抗药机制研究奠定基础。

The emergence and spread of drug-resistant Mycobacterium tuberculosis, especially multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) strains, is an important cause of the increased incidence of tuberculosis. Therefore, the discovery of new therapeutic drugs is an important research direction of current tuberculosis treatment. As a target of various antibiotics, it is of great significance for the functional study of mycobacterial ribosomes. Pathogenic M. tuberculosis grows slowly and culture conditions are harsh. It is often replaced by fast-growing, non-pathogenic Mycolicibacterium smegmatis in functional studies. However, there are significant differences in the number of ribosomes and metabolic diversity between these two species. Is there an intrinsic link between ribosome, metabolism, and growth rate? To explore such issues，we have adopted a strategy of synthetic biology：learning by building. We have assembled the ribosomal genes distributed in 24 locations of the M. tuberculosis genome into one gene cluster, and intend to integrate them into the chromosome of M. smegmatis, then carry out the systematic Knockout of endogenous ribosomes genes. By replacing the M. smegmatis ribosome genes, we will explore the relationship between ribosome gene copy number and growth rate, gene chromosome arrangement and express regulation, ribosome heterogeneity and translational regulation. Some functionally important ribosome genes may be identified by the mutants screening, and the study may lead to the construction of a series of M. smegmatis mutants in which most or all of the ribosomal genes have been replaced by M. tuberculosis orthologs. These mutants will be the new chassis for future drug screening or ribosome resistance studies.