GENE CLONING

 

1) Phage - Vector that transduce bacterial DNA from cell to another.

                - Infect cells more efficient than plasmid transformed cells, higher yield of clones.

                - About 1/3 of 𝛌 genome is not essential, can be replaced with foreign DNA.

 

2) Cosmid- Plasmid with 𝛌 contain phage cos site (cohesive end + 𝛌)

                  - Cohesive end are essential for packaging DNA into phage heads.

 

3) Phagemid- Generally small plasmid

                      - Have ability to accept larger DNA insert.

 

4) Bacteria Artificial Chromosome (BAC)

    - Able to carry ~ 20kb of inserted DNA sequences.

    - Contain selectable antibiotic marker ( ) and lac Z.

    - Used in genomic project to sequence the genes.

 

5) Yeast Artificial Chromosome (YAC)

     - Allow cloning of foreign genomic DNA in yeast cells.

     - Able to get eukaryotic protein product with posttranslational modification.

 

6) Expression vector

     - Allow expression of cloned gene to gene product, usually protein.

     - Used to isolate specific cDNAs.

     - Used in transformation to generate transgenic plants, animals, microbes

 

7) Ti plasmid

    - Large, circular double stranded DNA molecule with ~200kb

    - Integrate into plant genome, stably transmitted through mitosis and meiosis.