Chapter 15
Cloning Vectors
ANNETTE DEICHMANN AND KLAUS DEICHMANN
lntroduction
For investigation of the sequence, structure and function ofDNA fragments
a large number of identical copies is required. The production of large
amounts of DNA from a small number of copies is called cloning (Levin
1985; Winnacker 1985). For cloning, one can make use ofthe cell's replication mechanisms. A question arises, which is how to transport a DNA fragment into the cell and how to initiate its replication. Forthis purpose, cloning vectors have been designed. These vectors are descendants of naturallyoccuring, autonomaus DNA-species in bacterial cells - the bacteriophage
and plasmids. Insertion of foreign DNA fragments into the genomes of
phages and plasmids, produces a piece of recombinant DNA called a hybrid
or a chimera. The ability of bacteriophage and plasmids to be amplified in
bacterial cells is not alteredas long as the inserted DNA does not exceed a
certain size. The inserted DNA sequences does not disrupt a vector sequence that is essential for its function. Furthermore, there should be a
growth advantage for bacterial cells containing these hybrids and easy selection of chimerk vectors over native vectors. In short, vectors should provide us with:
• Simple methods for multiplication of specific DNA fragments
• High reliability regarding the identity of the copies
• Simple sequencing procedures tobe carried out directly on the recombinant vector
Annette Deichmann, Correspondence to Klaus Deichmann, Universitäts-Kinderklinik,
Mathildenstrasse 1, Freiburg, 79106, Germany (phone +49-7612704300; fax +497612706372; e-mail deichman@kkl200.ukl.uni-freiburg.de)
Cloning Vectors
ANNETTE DEICHMANN AND KLAUS DEICHMANN
lntroduction
For investigation of the sequence, structure and function ofDNA fragments
a large number of identical copies is required. The production of large
amounts of DNA from a small number of copies is called cloning (Levin
1985; Winnacker 1985). For cloning, one can make use ofthe cell's replication mechanisms. A question arises, which is how to transport a DNA fragment into the cell and how to initiate its replication. Forthis purpose, cloning vectors have been designed. These vectors are descendants of naturallyoccuring, autonomaus DNA-species in bacterial cells - the bacteriophage
and plasmids. Insertion of foreign DNA fragments into the genomes of
phages and plasmids, produces a piece of recombinant DNA called a hybrid
or a chimera. The ability of bacteriophage and plasmids to be amplified in
bacterial cells is not alteredas long as the inserted DNA does not exceed a
certain size. The inserted DNA sequences does not disrupt a vector sequence that is essential for its function. Furthermore, there should be a
growth advantage for bacterial cells containing these hybrids and easy selection of chimerk vectors over native vectors. In short, vectors should provide us with:
• Simple methods for multiplication of specific DNA fragments
• High reliability regarding the identity of the copies
• Simple sequencing procedures tobe carried out directly on the recombinant vector
Annette Deichmann, Correspondence to Klaus Deichmann, Universitäts-Kinderklinik,
Mathildenstrasse 1, Freiburg, 79106, Germany (phone +49-7612704300; fax +497612706372; e-mail deichman@kkl200.ukl.uni-freiburg.de)