Restriction enzymes: nomenclature, methods of recognizing and cutting DNA, how to make a restriction map.
Other enzymes used for cloning: polymerase, ligase, clonase, topoisomerase et al.
The vectors used for cloning of DNA: plasmids, viral vectors, cosmids, BACs, YACs.
Different methods of DNA cloning: cloning bay using restriction enzymes, TA cloning, TOPO TA cloning, Gateway cloning.
Libraries: gene and genome library, differences in preparation methodologies.
Identification and selection of certain specific DNA molecules from a library of genes.
Analysis of gene: analysis of gene expression by Northern hybridization, reverse transcription (cDNA synthesis from mRNA) and realtime PCR.
Expression of recombinant proteins: the selection of the host and vector (bacteria, insect cells, etc.).
The basic principles of gene cloning into expression vectors. Development of primers for the cloning and modification of genes.
Purification of the recombinant protein by affinity chromatography and detection of proteins by SDS PAGE and Western blot hybridization.
How to analyze the interaction between proteins. Pull down assay and yeast two hybrid system.
1.Understand how we can manipulate DNA and RNA by using different enzymes and methods to create recombinant molecules
2.Understand methods for gene and protein analysis, and how to applied them in biotechnology.
3.Acquire insight into ways to manipulate genes in vitro and know how manipulate existing properties, and how to obtain new properties in transgenic organisms
4.Understand how gene manipulation help us to understand the function of specific genes and specific biological process.