Faculty of Science

Objectives

To present an in-depth treatment of recombinant DNA technology as the foundation of modern biotechnology and to show how its tools can be employed in the guided production of goods and services.

Contents

What is Biotechnology? The scope of Biotechnology!

Principles and methods of recombinant DNA technology- Restriction modification enzymes used in recombinant DNA technology. Hybridization, cloning, sequencing, polymerase chain reaction; gene manipulations; cloning vectors, cloning in E.coli, plasmids, bacteriophages and cosmid vectors, cloning strategies, genomic and cDNA library; Screening of gene libraries – screening by DNA hybridization, immunological assay and protein activity; DNA delivery methods - physical methods and biological methods; expression of cloned genes in E. coli, products made in E. coli by genetic engineering; cloning in yeast: transformation in yeast, yeast vector development; features of yeast promoter and expression of cloned genes; yeast 2-hybrid system; plasmid shuffling to explore interactive domains of multimeric proteins; the cassette model for mating type switches and silencing of genes. Strong and regulatable promoters; increasing protein production; Fusion proteins; Translation expression vectors; DNA integration into bacterial genome; Increasing secretions; Metabolic load, Directed mutagenesis; transposon mutagenesis, Gene targeting, Site specific recombination.