Transmission of genetic material in bacteria, Conjugation,and gene recombination in E.

coli, Transduction and Transformation

SUBMITTED TO:-

Dr. Zahra
SUBMITTED BY:-

Uzma Mushtaq
ROLL NO. 9

University of Education Bank Road Campus Lahore

Some Organisms Transmit Genetic Material to Offspring without Cell Division

Prokaryotes, which include bacteria and single-celled microorganisms called Archaea, usually pass their chromosomal DNA on to their offspring asexually. In other words, a bacterial cell reproduces by simply replicating its chromosome and dividing into two daughter cells. The daughter cells that result from this division are genetically identical to each other and to the original parent cell. Thus, over time, asexual reproduction in

bacteria can lead to a population of hundreds of thousands of cells, all of which are genetically identical to a lone original parent cell. Given their asexual method of reproduction, it is tempting to think that bacteria are sorely lacking in genetic variation, but this is not the case. Prokaryotic cells have developed a number of methods for recombining their genetic material, which, in turn, contributes to their genetic diversity. The three most common ways that bacteria diversify their DNA are transformation, conjugation, and transduction. However, not all types of bacterial cells are capable of engaging in all three processes.

Transformation
Transformation is a process by which a susceptible or "competent" bacterial cell acquires new genetic material from its environment. There are two types of transformation: natural and artificial. But where does the environmental DNA required for transformation come from? And how does this DNA become part of a bacterium's genome? Natural transformation, as its name implies, is a natural mechanism used by some bacterial cells to take up DNA from the environment. This environmental DNA was, at one point, located in other bacteria. For instance, when bacteria die and disintegrate, their chromosomal DNA is released. Fragments of this DNA remain in the environment and are freely available to other living cells, including other bacteria. These naturally occurring DNA fragments can enter a living bacterium through its cell membrane, after contact with that membrane. If the DNA is double stranded, one of the strands will pass across the cell membrane into the cell, and the other strand will be dissolved, or hydrolyzed. Parts of the newly introduced single-stranded DNA molecule may then recombine with similar regions on the bacterial chromosome and become incorporated into the bacterium's genome. In contrast, during artificial transformation, DNA uptake by bacterial host cells occurs under certain laboratory conditions. In the lab, scientists often introduce foreign DNA into bacterial cells via transformation in order to study specific genes and their functions. Typically, these researchers use E. coli cells that have been chemically treated so that their outer cell membranes are permeable to foreign DNA. In addition, transformation can be induced by electroporation, a process in which the bacterial host cells are subjected to an electric field that allows molecules to pass more easily across the membrane. Heat shock is another way that transformation can occur, wherein host cells are exposed to extreme temperatures that also cause the cell membrane to temporarily allow molecules of foreign DNA into the cell. Within the lab environment, bacteria are also commonly transformed with sequences of DNA called plasmid vectors. These naturally occurring DNA molecules are circular, and they can replicate inside a bacterium independent of the bacterial chromosome (which can also be circular). Plasmid vectors can be used to clone, transfer, and manipulate genes. Often, these plasmids carry a gene for antibiotic resistance, which means that

researchers can select for cells that are resistant to a given antibiotic in order to determine whether a bacterium has been successfully transformed. The following animation depicts the process of transformation:

Conjugation
Conjugation is a process by which one bacterium transfers genetic material to another bacterium through direct contact. During conjugation, one of the bacterial cells serves as the donor of the genetic material, and the other serves as the recipient. The donor bacterium carries a circular, double-stranded DNA sequence called the fertility factor, or F-factor. The F-factor allows the donor to produce a thin, tubelike protuberance called a pilus. The donor uses the pilus to contact the recipient. The pilus then shortens and draws the two bacteria together, at which time the donor bacterium transfers genetic material to the recipient bacterium. This genetic material is in the form of a plasmid, or a small, circular piece of non-chromosomal DNA. The newly transferred genetic material often provides the recipient bacterium with some sort of genetic advantage. In many cases, conjugation results in the transfer of a plasmid containing an antibiotic resistance gene. The following animation depicts two bacteria exchanging DNA via conjugation:

Transduction
Finally, transduction is a process by which a virus transfers genetic material from one bacterium to another bacterium. This process depends on a specific type of virus called a bacteriophage, which is capable of infecting bacterial cells and using them as hosts to produce more viruses. At the beginning of a transduction cycle, a bacteriophage injects its DNA into a host bacterium. The phage DNA then takes over the host cell's machinery and directs it to synthesize and assemble more phages. During this process, the host cell's DNA breaks into fragments; after that, the host cell replicates the phage DNA and assembles new phages. Occasionally, some of the bacterial host cell's DNA is included with the phage DNA during assembly. Once phage assembly is complete, the bacterial cell breaks open, and the newly assembled phages are released into the environment. This is called a lytic cycle because the original bacterial host cell is destroyed, or lysed. Later, when one of the newly released bacteriophages infects a new bacterium, any bacterial DNA that the phage contains may become incorporated into the genome of the new host. The original bacterial host cell is not always destroyed during transduction, however. In some cases, the phage DNA does not direct the host cell to produce more phages; instead, it incorporates itself into the chromosomal DNA of the bacterial host cell. This is called a lysogenic cycle. The phage DNA is then maintained within the bacterial chromosome through many generations of cell division. Eventually, at a point in the future when

conditions are right, the phage DNA removes itself from the bacterial chromosome and initiates a lytic cycle. The following animation shows the process of transduction. The first part of the animation depicts a lytic cycle, and the second part shows a lysogenic cycle:

Further Exploration Key Questions How is DNA organized in prokaryotes? Key Concepts prokaryote | plasmid

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Transformation : transfer of genetic materials without any physical contact or mediators. The genetic materials can be from other bacteria or even other organisms. The genetic materials come into the bacterium through specific transporter on the cell

membrane. Transduction : transfer of genetic materials through mediators, usually virus, especially bacteriophage (virus that infect bacteria). The genetic materials from other bacteria that are infected by the virus, can be accidentally packaged inside the bacteriophage head when the viral particles are assembled prior to release from infected bacteria. There virus can infect other bacteria, but instead of transferring viral gene, it transfers bacterial gene into the bacteria. Conjugation : transfer of genetic materials among bacteria through physical contact using a structure called pilus. Not all bacteria can transfer their genetic materials. Only those who have the so-called F-plasmid can transfer this plasmid to other bacteria. The recipient will become F positive bacteria and are able to transfer the plasmid to other socalled F minus bacteria. There other kinds of conjugation, this is called Hfr (high frequency recombination) with different mode of transfer, but similarly require pilus for transfer. This kind of conjugation is used by scientist to map bacterial genome. After transfer, the DNA molecules can exist in two forms, either as DNA molecules separate from the bacterial chromosome (an episome), or can become part of the bacterial chromosome. The study of basic mechanisms used by bacteria to exchange genes allowed scientists to develop many of the essential tools of modern molecular biology. Bacterial conjugation refers to the transfer of DNA between bacterial cells that requires cell-to-cell contact. Joshua Lederberg and Edward Tatum first

A laboratory technician performing an Analytical Profile Index (API) test on bacteria. described conjugation in 1946 when they discovered the F factor (an episome) that can move between Escherichia coli cells. The F factor is one of the most well studied conjugative plasmids (plasmids are circular episomes) and is the most well studied conjugative system. There are many different conjugal plasmids carried by members of most bacterial species. Conjugal plasmids that carry antibiotic resistance genes are called R factors. The F factor and R factors usually exist as episomes and each carries functions that allow it to replicate its DNA and thus be inherited by the daughter cells after binary fission. However, conjugative plasmids also express transfer functions that allow the movement of DNA from a donor to a recipient cell; this is the process of conjugation. The steps of bacterial conjugation are: mating pair formation, conjugal DNA synthesis, DNA transfer, and maturation. The main structure of the F factor that allows mating pair formation is the F pilus or sex pilus (a long thin fiber that extends from the bacterial cell surface). There are one to three pili expressed on an E. coli cell that carries the F factor, and one pilus will specifically interact with several molecules on the recipient cell surface (attachment). About twenty genes on the F factor are required to produce a functional pilus, but the structure is mainly made up of one protein , pilin. To bring the donor and recipient cell into close proximity, the F pilus retracts into the donor cell by removing pilin protein monomers from the base of the pilus to draw the bacterial cells together. Once a stable mating pair is formed, a specialized form of DNA replication starts. Conjugal DNA synthesis produces a single-stranded copy of the F factor DNA (as opposed to a double-stranded DNA that is formed by normal replication). This DNA

strand is transferred into the recipient cell. Once in the recipient cell, the single-stranded copy of the F plasmid DNA is copied to make a double-stranded DNA molecule, which then forms a mature circular plasmid. At the end of conjugation the mating pair is broken and both the donor and the recipient cells carry an identical episomal copy of the F factor. All of the approximately one hundred genes carried on the F factor can now be expressed by the recipient cell and will be inherited by its offspring. In addition to transferring itself, the F factor can also transfer chromosomal genes between a donor and recipient cell. The F factor can be found inserted (integrated) into the bacterial chromosome at many locations in a small fraction of bacterial cells. An integrated F factor is replicated along with the rest of the chromosome and inherited by offspring along with the rest of the chromosome. When a mating pair is formed between the donor cell carrying an integrated F factor and a recipient cell, DNA transfer occurs as it does for the episomal F factor, but now the chromosomal sequences adjacent to the integrated F factor are transferred into the recipient. Since these DNA sequences encode bacterial genes, they can recombine with the same genes in the recipient. If the donor gene has minor changes in DNA sequence from the recipient gene, the different sequence can be incorporated into the recipient gene and inherited by the recipient cell's offspring. Donor cells that have an integrated copy of the F factor are called Hfr strains (High frequency of recombination).

Transduction
The second way that DNA is transferred between bacterial cells is through a phage particle in the process of transduction. Joshua Lederberg and Norton Zinder first discovered transduction in 1956. When phage inject their DNA into a recipient cell, a process occurs that produces new bacteriophage particles and kills the host cell (lytic growth). Some phage do not always kill the host cell (temperate phage), but instead can be inherited by daughter host cells. Therefore acquisition of a so-called temperate "prophage" by a recipient cell is a form of transduction. Many phage also have the ability to transfer chromosomal or plasmid genes between bacterial cells. During generalized transduction any gene can be transferred from a donor cell to a recipient cell. Generalized transducing phage are produced when a phage packages bacterial genes into its capsid (protein envelope) instead of its own DNA. When a phage particle carrying bacterial chromosomal genes attaches to a recipient cell, the DNA is injected into the cytoplasm where it can recombine with a homologous DNA sequences. Some bacteriophage can pick up a subset of chromosomal genes and transfer them to other bacteria. This process is called specialized transduction

Transduction

Bacterial conjugation. The bacterium on the left passes a copy of the F plasmid to the bacterium on the right, converting it from an F - cell to an F + cell. since only a limited set of chromosomal genes can be transferred between bacterial cells.

A scanning electron micrograph of bacterial DNA plasmids.

Transformation
The third main way that bacteria exchange DNA is called DNA transformation. Some bacteria have evolved systems that transport free DNA from the outside of the bacterial cell into the cytoplasm. These bacterial are called "naturally competent" for DNA transformation. Natural DNA transformation of Streptococcus pneumonaiae provided the first proof that DNA encoded the genetic material in experiments by Oswald Avery and colleagues. Some other naturally competent bacteria include Bacillus subtilis, Haemophilus influenzae, and Neisseria gonorrhoeae. Other bacterial species such as E. coli are not naturally competent for DNA transformation. Scientists have devised many ways to physically or chemically force noncompetant bacteria to take up DNA. These methods of artificial DNA transformation form the basis of plasmid cloning in molecular biology. Most naturally competent bacteria regulate transformation competence so that they only take up DNA into their cells when there is a high density of cells in the environment. The ability to sense how many other cells are in an area is called quorum sensing. Bacteria that are naturally competent for DNA transformation express ten to twenty proteins that form a structure that spans the bacterial cell envelope. In some bacteria this structure also is required to form a particular type of pilus different than the F factor pilus. Other bacteria express similar structures that are involved in secreting proteins into the exterior medium (Type II secretion). Therefore, it appears that DNA transformation and protein secretion have evolved together. During natural DNA transformation, doubled-stranded DNA is bound to the recipient cell surface by a protein receptor. One strand of the DNA is transported through the cell envelope, where it can recombine with similar sequences present in the recipient cell. If the DNA taken up is not homologous to genes already present in the cell, the DNA is usually broken down and the nucleotides released are used to synthesize new DNA during normal replication. This observation has led to the speculation that DNA transformation competence may have originally evolved to allow the acquisition of nucleic acids for food. The source of DNA for transformation is thought to be DNA released from other cells in the same population. Most naturally competent bacteria spontaneously break apart by expressing enzymes that break the cell wall. Autolysis will release the genomic DNA into the environment where it will be available for DNA transformation. Of course, this results in the death of some cells in the population, but usually not large numbers of cells. It appears that losing a few cells from the population is counterbalanced by having the possibility of gaining new traits by DNA transformation.

Some Organisms Transmit Genetic Material to Offspring without Cell Division

Prokaryotes, which include bacteria and single-celled microorganisms called Archaea, usually pass their chromosomal DNA on to their offspring asexually. In other words, a bacterial cell reproduces by simply replicating its chromosome and dividing into two daughter cells. The daughter cells that result from this division are genetically identical to each other and to the original parent cell. Thus, over time, asexual reproduction in bacteria can lead to a population of hundreds of thousands of cells, all of which are genetically identical to a lone original parent cell. Given their asexual method of reproduction, it is tempting to think that bacteria are sorely lacking in genetic variation, but this is not the case. Prokaryotic cells have developed a number of methods for recombining their genetic material, which, in turn, contributes to their genetic diversity. The three most common ways that bacteria diversify their DNA are transformation, conjugation, and transduction. However, not all types of bacterial cells are capable of engaging in all three processes.