Plasmid

Plasmids are circular DNA molecules that are separate from the chromosomal DNA (Fig. 1). They usually occur in bacteria, sometimes in eukaryotic organisms (e.g., the 2-micron-ring in Saccharomyces cereviesiae). Their size varies from 1 to 250 kbp or thousand base pairs. There are from one copy, for large plasmids, to fifty copies of the same plasmid present in a single cell.

http://www.nupedia.com/newsystem/upload_file/670/plasmids.png Description : This image shows a line drawing of a bacterium with its chromosomal DNA and several plasmids within it. The bacterium is drawn as a large oval. Within the bacterium, small to medium size circles illustrate the plasmids, and one long thin closed line that intersects itself repeatedly illustrates the chromosomal DNA.

Figure 1 : Schematic drawing of a bacterium with plasmids enclosed. (Image from Nupedia.) (1) Chromosomal DNA. (2) Plasmids.

Plasmids usually contain one or two genes that confer a selective advantage on the bacterium harboring them, e.g., the ability to build an antibiotic resistance. Every plasmid contains at least one DNA sequence that serves as an origin of replication or ori (a starting point for DNA replication), which enables the plasmid DNA to be duplicated independently from the chromosomal DNA (Fig. 2).

http://www.nupedia.com/newsystem/upload_file/670/example_plasmid.png Description : This image shows a line drawing of a plasmid. The plasmid is drawn as two concentric circles that are very close together, with two large segments and one small segment marked off on the two circumferences. The two large segments indicate antibiotic resistances and the small segment indicates an origin of replication.

Figure 2 : Schematic drawing of a plasmid with antibiotic resistances (1&2) and an ori(3). (Image from Nupedia.)

Episomes are plasmids that can integrate themselves into the chromosomal DNA of the host organism (Fig. 3). For this reason, they can stay intact for a long time, be duplicated with every cell division of the host, and become a basic part of its genetic makeup.

http://www.nupedia.com/newsystem/upload_file/670/plasmid_episome.png Description : This image shows a line drawing that compares the activity of non-integrating plasmids, on the top, with episomes, on the bottom, during cell division. The upper half of the image shows a bacterium with its chromosomal DNA and plasmids dividing into two identical bacteria, each with their chromosomal DNA and plasmids. The lower half of the image shows a bacterium with its chromosomal DNA, but with an episome. Next to this bacterium, we see the same bacterium, but after the episome has integrated into the chromosomal DNA and has become a part of it. This second bacterium now divides into two bacteria identical to it, each with an episome integrated into it.

Figure 3 : Comparison of non-integrating plasmids (top) and episomes (bottom). (Image from Nupedia.) (1) Chromosomal DNA. (2) Plasmids. (3) Cell division. (4) Chromosomal DNA with integrated plasmids.

There are two basic groups of plasmids, conjugative and non-conjugative. Conjugative plasmids contain a so-called tra-gene, which can initiate conjugation, the sexual exchange of plasmids, with another bacterium (Fig. 4). Non-conjugative plasmids are incapable of initiating conjugation, and therefore, their movement to another bacterium, but they can be transferred together with conjugative plasmids, during conjugation.

http://www.nupedia.com/newsystem/upload_file/670/conjugative_plasmids.png Description : This image is a line drawing of bacterial conjugation. The image shows, going from the top to the bottom, two bacteria before, during, and after conjugation. On the top then are two bacterium, before conjugation, each with their own chromosomal DNA. Only one bacterium shows a plasmid. In the middle, are the same two bacterium during conjugation. A pilius (connection) forms between the two bacteria and a linear copy of the plasmid is transported through the pilius to the other bacterium. On the bottom, are the same two bacterium after conjugation. The pilius is now gone and each bacterium has a plasmid.

Figure 4 : Schematic drawing of bacterial conjugation. (Image from Nupedia.) (1) Chromosomal DNA. (2) Plasmids. (3) Pilus.

Several different types of plasmids can coexist in a single cell, e.g., up to seven in E. coli. Two plasmids can be incompatible, resulting in the destruction of one of them. Therefore, plasmids can be assigned into incompatability groups, depending on their ability to coexist in a single cell.

An obvious way of classifying plasmids is by function. There are five main classes:

• Fertility-(F-)plasmids, which contain only tra-genes. Their only function is to initiate conjugation.
• Resistance-(R-)plasmids, which contain genes that can build a resistance against antibiotics or poisons.
• Col-plasmids, which contain genes that code for (determine the production of) colicines, proteins that can kill other bacteria.
• Degrative plasmids, which enable the digestion of unusual substances, e.g., toluole or salicylic acid.
• Virulence plasmids, which turn the bacterium into a pathogen.

Plasmids that exist only as a single copy in each bacterium are, upon cell division, in danger of being lost in one of the segregating bacteria. To ensure that the cell has an "interest" in keeping a copy of the plasmid in each dividing cell, some plasmids include an addiction system. They produce both a long-lived poison and its short-lived antidote. The cell that keeps a copy of the plasmid will survive, while the cell without the plasmid will die because it is running out of antidote shortly.

Plasmids serve as important tools in genetics and biochemical labs, where they are commonly used to multiply or express particular genes. There are many plasmids that are commercially available for such uses. Initially, the gene to be replicated is inserted in a plasmid. But, these plasmids contain, in addition to the inserted gene, one or more genes with antibiotic resistance. The plasmids are next inserted into bacteria, which are then grown on specific antibiotic(s). As a result, only the bacteria with antibiotic resistance can survive, the very same bacteria containing the genes to be replicated. The antibiotic(s) will, however, kill those bacteria that did not receive a plasmid, because they have no antibiotic resistance genes. In this way the antibiotic(s) acts as a filter selecting out only the modified bacteria. This is a cheap and easy way of mass-producing a gene or the protein it codes for--for example, insulin or even antibiotics.