Genetic Engineering an der Hochschule Bonn-Rhein-Sieg

Any artificially produced DNA-molecule which brings together DNA sequences that are not usually found together in nature. 

Techniques and procedures that are necessary to produce recombinant DNA and to transfer it into living cells.

In 1972.

Direct evolution is a speed up evolution in a test tube and involves the following steps: 

1. DNA isolation 

2. Introduction of random mutagenesis 

3. Repeat 1 and 2

4. Transfer back to organism

5. Analysis

Earlier, UV light was used for dimer formation.

Today, we use error-prone PCR.

generation and propagation of recombinant DNA.

1. DNA isolation of vector and insert (gene of interest) 

2. Process DNA with restriction enzymes 

3. Ligation 

4. Transformation of host cell 

5. Selection and Screening 

When recombinant DNA was introduced into it. 

- to produce useful recombinant molecules

- to improve agronomic traits (by overexpression/repression of certain proteins or by showing new traits leading to e.g. herbicide resistance) 

Advantages:

- genetics well understood

- unlimited quantities of protein generation possible 

- fermentation technology well understood (faster growth, cheap medium) 

Disadvantages: 

- eukaryotic proteins cannot be folded correctly by prokaryot and might be inactive

- formation of inclusion bodies possible

- Growth of cells (e.g. can they grow in fermenter?)

- Cell lines may have mutations and their extensiveness is not known (e.g. are they free of endogenous viruses?) 

Modified animals that can produce desired products.