Cloning, of DNA

Regardless of the strategy one uses to identify and clone the gene of interest, these genetic clones of DNA encoding for... 

Cloning of DNA has not only provided an essential step in sequence determination but also has given birth to a new industry devoted to producing proteins from genes cloned in bacteria, yeast, or other cells. Human... 

Construction of the gene-bearing vector. Once the clone of DNA has been isolated, this is put into a vector (in this case a plasmid) which will transfer it from the host to the recipient organism. Again, by the use of restriction enzymes to cut the plasmid vector and the cloned DNA, the cloned DNA can now be inserted into the vector. 

Directional cloning of DNA fragments at a large distance from an initial probe ... 

One of the limitations of screening environmental isolates is the cultivation of the organisms. In a single gram of soil, more than 108 microbes may be present, the majority of which cannot be cultivated. It is clear from molecular ecological studies that traditional cultivation techniques only capture a fraction, usually less than 1 percent, of the available diversity. To overcome this, direct cloning of DNA from environmental samples enables an unbiased representation of the microbial... 

Figure 2. Schematic of insertion of foreign DNA fragment into plasmid vector and cloning of DNA molecules in host cells.

Polli JW, Kincaid RL (1992) Molecular cloning of DNA encoding a calmodulin-dependent phosphodiesterase enriched in striatum. Proc Natl Acad Sci USA 59 11079-11083. 

The cosmid is another variation of the plasmid and viral DNA vectors that has been developed to allow the cloning of DNA fragments that range from 40 to 50 kb. As the linear bacteriophage A DNA is... 

Multiple cloning sites (MCS). These are fragments of DNA that contain a number of unique different restriction sites, enabling the use of a choice of restriction enzymes for the cloning of DNA fragments. 

Fig. 24.1 Simplified diagram of the plasmid pUCl 8. lacZ represents the insertional inactivation marker coding for fl-galactosidase activity. A multiple cloning site (MCS) is present within the LacZ gene to enable the cloning of DNA fragments. Ori represents the origin of replication which, in this case, works in Escherichia coli. Finally, Ampr represents an ampicillin resistance marker.

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