Abstract

DNA cloning and the analysis of plasmid structure and function are fundamental techniques in molecular biology, enabling the detailed study and manipulation of genetic material. This article provides a comprehensive overview of the methods and applications associated with plasmid-based DNA cloning. Plasmids, which are small, circular, double-stranded DNA molecules found in bacteria, serve as versatile vectors for the replication and expression of foreign genes. The process of cloning involves the insertion of target DNA sequences into plasmid vectors using restriction enzymes and DNA ligases, followed by their introduction into host cells via transformation. Subsequent analysis of plasmid structure—including restriction mapping, sequencing, and gel electrophoresis—allows researchers to verify successful cloning and investigate gene expression, regulatory elements, and functional domains. The study further explores advanced applications such as recombinant protein production, gene editing, and synthetic biology. Emphasis is also placed on the design of expression vectors, selectable markers, and reporter genes used in functional assays. Overall, this article highlights the pivotal role of plasmid-based cloning in genetic engineering, biomedical research, and biotechnology, offering insights into both foundational concepts and modern innovations in plasmid analysis and manipulation.