Molecular genetic engineering

Apuntes

•Main Themes The text offers an overview of molecular genetic engineering techniques, focusing on DNA transfer methods and cloning vectors. •Transformation and Transfection - Transformation involves competent cells absorbing DNA through binding, uptake, and RecA-mediated homologous recombination. - Transfection specifically refers to bacterial transformation using bacteriophage DNA. •Conjugation - Bacterial conjugation transfers DNA via conjugative plasmids, mainly the F plasmid, which encodes replication, transfer (tra), and transposable elements. - The process requires a sex pilus; DNA transfer occurs through rolling circle replication, with the pilus retracting to establish contact and transfer a nicked single strand that replicates in the recipient. •Transduction - Transduction transfers DNA via bacteriophages, with generalized (random DNA) and specialized (specific genes) types. - Generalized transduction is less efficient; specialized transduction results from incorrect prophage excision, carrying adjacent host genes. - Lysogenization can lead to phage conversion, altering host phenotypes, as seen in Salmonella enterica and Corynebacterium diphtheriae. •Cl...