Recombination

Cutting and covalent joining of DNA sequences.

Inter-molecular recombination

Intra-molecular recombination

Between two different DNA molecules

Between two regions of a single DNA molecule

Recombination may occur between linear and circular ds DNA templates or between two circular ds DNA templates.

Intermolecular recombination
Donor dsDNA : Red Recipient dsDNA: Black Recombinant DNA: Red and Black line. X: Reciprocal cross-over between the two dsDNA molecules. half-X : Non- reciprocal cross-over between the two dsDNA molecules.

Case 1 ::

(A): Single, reciprocal cross-over between two linear dsDNA (B): Single, non- reciprocal cross-over between two linear dsDNA

Case 2 ::

(C): Double, reciprocal cross-over between two linear dsDNA

(D): Double, non- reciprocal cross-over between two linear dsDNA

Case 3 ::

(E): Single reciprocal cross-over between linear dsDNA & circular dsDNA

Case 4 ::

(F): Double, reciprocal cross-over Single between linear dsDNA & circular dsDNA (G): Double, Non- reciprocal cross-over Single between linear dsDNA & circular dsDNA

Intramolecular recombination

Recombination between two regions on a single dsDNA molecule. Looping out of the intermediate region. Products :: A shorter dsDNA molecule and a separate circular dsDNA molecule. Reverse reaction can also occur - resulting in the integration of a circular dsDNA molecule into another dsDNA molecule

Types of recombination

Site-specific recombination

Homologous recombination

DNA strand exchange takes place between segments possessing only a limited degree of sequence homology. Occurs at specific DNA sequences. Specific sequences are recognized by enzymes and catalyze recombination with a specific recipient DNA.

Occurs between DNA sequences that are homologous -- that is, the two DNA sequences are nearly identical.

Non-Homologous recombination

Random insertion into or replacement of DNA sequences of target sequence.

Homologous Recombination from a Molecular Perspective

Site-specific Recombination
DNA strand exchange takes place between segments possessing only a limited degree of sequence homology. Occurs at specific DNA sequences (recombination sites). Specific sequences are recognized by enzymes (recombinases), and catalyze recombination with a specific recipient DNA.

Integrase in :: Recombinase attP site in & attB site on E.coli::

Specific sites of recombination.
Integrase cleaves DNA at both the the specific sequences & insert the phage DNA into the bacterial genome between gal & bio operons.

Two hybrid sites are formed as a result:- attL & attR

Cre-Lox recombination Cre protein:: Recombinase loxP site:: Specific site of recombination
When cells that have loxP sites in their genome express Cre, a recombination event can occur between the loxP sites. The double stranded DNA is cut at both loxP sites by the Cre protein. The strands are then rejoined with DNA ligase.

The result of recombination depends on the orientation of the loxP sites. (A) For two lox sites on the same chromosome arm, inverted loxP sites will cause an inversion of the intervening DNA. (A) If loxP sites are on different chromosomes translocation occurs. (A) Direct repeat of loxP sites on same chromosome will cause a deletion event of intervening DNA.