Transduction (genetics)

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Transduction Transduction image.pdf
Transduction
This is an illustration of the difference between generalized transduction, which is the process of transferring any bacterial gene to a second bacterium through a bacteriophage and specialized transduction, which is the process of moving restricted bacterial genes to a recipient bacterium. While generalized transduction can occur randomly and more easily, specialized transduction depends on the location of the genes on the chromosome and the incorrect excision of the a prophage. Transduction illustration.pdf
This is an illustration of the difference between generalized transduction, which is the process of transferring any bacterial gene to a second bacterium through a bacteriophage and specialized transduction, which is the process of moving restricted bacterial genes to a recipient bacterium. While generalized transduction can occur randomly and more easily, specialized transduction depends on the location of the genes on the chromosome and the incorrect excision of the a prophage.

Transduction is the process by which foreign DNA is introduced into a cell by a virus or viral vector. [1] An example is the viral transfer of DNA from one bacterium to another and hence an example of horizontal gene transfer. [2] Transduction does not require physical contact between the cell donating the DNA and the cell receiving the DNA (which occurs in conjugation), and it is DNase resistant (transformation is susceptible to DNase). Transduction is a common tool used by molecular biologists to stably introduce a foreign gene into a host cell's genome (both bacterial and mammalian cells).

Contents

Discovery (bacterial transduction)

Transduction was discovered in Salmonella by Norton Zinder and Joshua Lederberg at the University of Wisconsin–Madison in 1952. [3]

In the lytic and lysogenic cycles

Transduction happens through either the lytic cycle or the lysogenic cycle. When bacteriophages (viruses that infect bacteria) that are lytic infect bacterial cells, they harness the replicational, transcriptional, and translation machinery of the host bacterial cell to make new viral particles (virions). The new phage particles are then released by lysis of the host. In the lysogenic cycle, the phage chromosome is integrated as a prophage into the bacterial chromosome, where it can stay dormant for extended periods of time. If the prophage is induced (by UV light for example), the phage genome is excised from the bacterial chromosome and initiates the lytic cycle, which culminates in lysis of the cell and the release of phage particles. Generalized transduction (see below) occurs in both cycles during the lytic stage, while specialized transduction (see below) occurs when a prophage is excised in the lysogenic cycle.[ citation needed ]

As a method for transferring genetic material

Transduction by bacteriophages

The packaging of bacteriophage DNA into phage capsids has low fidelity. Small pieces of bacterial DNA may be packaged into the bacteriophage particles. There are two ways that this can lead to transduction.[ citation needed ]

Generalized transduction

Generalized transduction occurs when random pieces of bacterial DNA are packaged into a phage. It happens when a phage is in the lytic stage, at the moment that the viral DNA is packaged into phage heads. If the virus replicates using 'headful packaging', it attempts to fill the head with genetic material. If the viral genome results in spare capacity, viral packaging mechanisms may incorporate bacterial genetic material into the new virion. Alternatively, generalized transduction may occur via recombination. Generalized transduction is a rare event and occurs on the order of 1 phage in 11,000.

The new virus capsule that contains part bacterial DNA then infects another bacterial cell. When the bacterial DNA packaged into the virus is inserted into the recipient cell three things can happen to it:[ citation needed ]

  1. The DNA is recycled for spare parts.
  2. If the DNA was originally a plasmid, it will re-circularize inside the new cell and become a plasmid again.
  3. If the new DNA matches with a homologous region of the recipient cell's chromosome, it will exchange DNA material similar to the actions in bacterial recombination.

Specialized transduction

Specialized transduction is the process by which a restricted set of bacterial genes is transferred to another bacterium. Those genes that are located adjacent to the prophage are transferred due to improper excision. Specialized transduction occurs when a prophage excises imprecisely from the chromosome so that bacterial genes lying adjacent to it are included in the excised DNA. The excised DNA along with the viral DNA is then packaged into a new virus particle, which is then delivered to a new bacterium when the phage attacks new bacterium. Here, the donor genes can be inserted into the recipient chromosome or remain in the cytoplasm, depending on the nature of the bacteriophage.[ citation needed ]

When the partially encapsulated phage material infects another cell and becomes a prophage, the partially coded prophage DNA is called a "heterogenote".[ citation needed ]

An example of specialized transduction is λ phage in Escherichia coli . [4]

Lateral transduction

Lateral transduction is the process by which very long fragments of bacterial DNA are transferred to another bacterium. So far, this form of transduction has been only described in Staphylococcus aureus, but it can transfer more genes and at higher frequencies than generalized and specialized transduction. In lateral transduction, the prophage starts its replication in situ before excision in a process that leads to replication of the adjacent bacterial DNA. After which, packaging of the replicated phage from its pac site (located around the middle of the phage genome) and adjacent bacterial genes occurs in situ, to 105% of a phage genome size. Successive packaging after initiation from the original pac site leads to several kilobases of bacterial genes being packaged into new viral particles that are transferred to new bacterial strains. If the transferred genetic material in these transducing particles provides sufficient DNA for homologous recombination, the genetic material will be inserted into the recipient chromosome. Because multiple copies of the phage genome are produced during in situ replication, some of these replicated prophages excise normally (instead of being packaged in situ), producing normal infectious phages. [5]

Mammalian cell transduction with viral vectors

Rat nerve cells express red and green fluorescent proteins after viral transduction with two artificial adeno-associated viruses. Synapsin and CamK2 positive neurons.png
Rat nerve cells express red and green fluorescent proteins after viral transduction with two artificial adeno-associated viruses.

Transduction with viral vectors can be used to insert or modify genes in mammalian cells. It is often used as a tool in basic research and is actively researched as a potential means for gene therapy.[ citation needed ]

Process

In these cases, a plasmid is constructed in which the genes to be transferred are flanked by viral sequences that are used by viral proteins to recognize and package the viral genome into viral particles. This plasmid is inserted (usually by transfection) into a producer cell together with other plasmids (DNA constructs) that carry the viral genes required for the formation of infectious virions. In these producer cells, the viral proteins expressed by these packaging constructs bind the sequences on the DNA/RNA (depending on the type of viral vector) to be transferred and insert it into viral particles. For safety, none of the plasmids used contains all the sequences required for virus formation, so that simultaneous transfection of multiple plasmids is required to get infectious virions. Moreover, only the plasmid carrying the sequences to be transferred contains signals that allow the genetic materials to be packaged in virions so that none of the genes encoding viral proteins are packaged. Viruses collected from these cells are then applied to the cells to be altered. The initial stages of these infections mimic infection with natural viruses and lead to expression of the genes transferred and (in the case of lentivirus/retrovirus vectors) insertion of the DNA to be transferred into the cellular genome. However, since the transferred genetic material does not encode any of the viral genes, these infections do not generate new viruses (the viruses are "replication-deficient").[ citation needed ]

Some enhancers have been used to improve transduction efficiency such as polybrene, protamine sulfate, retronectin, and DEAE Dextran. [6]

Medical applications

See also

Related Research Articles

<span class="mw-page-title-main">Bacteriophage</span> Virus that infects and replicates within bacteria

A bacteriophage, also known informally as a phage, is a virus that infects and replicates within bacteria and archaea. The term was derived from "bacteria" and the Greek φαγεῖν, meaning "to devour". Bacteriophages are composed of proteins that encapsulate a DNA or RNA genome, and may have structures that are either simple or elaborate. Their genomes may encode as few as four genes and as many as hundreds of genes. Phages replicate within the bacterium following the injection of their genome into its cytoplasm.

<span class="mw-page-title-main">Lambda phage</span> Bacteriophage that infects Escherichia coli

Enterobacteria phage λ is a bacterial virus, or bacteriophage, that infects the bacterial species Escherichia coli. It was discovered by Esther Lederberg in 1950. The wild type of this virus has a temperate life cycle that allows it to either reside within the genome of its host through lysogeny or enter into a lytic phase, during which it kills and lyses the cell to produce offspring. Lambda strains, mutated at specific sites, are unable to lysogenize cells; instead, they grow and enter the lytic cycle after superinfecting an already lysogenized cell.

A provirus is a virus genome that is integrated into the DNA of a host cell. In the case of bacterial viruses (bacteriophages), proviruses are often referred to as prophages. However, proviruses are distinctly different from prophages and these terms should not be used interchangeably. Unlike prophages, proviruses do not excise themselves from the host genome when the host cell is stressed.

<span class="mw-page-title-main">Cloning vector</span> Small piece of maintainable DNA

A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The vector contains features that allow for the convenient insertion of a DNA fragment into the vector or its removal from the vector, for example through the presence of restriction sites. The vector and the foreign DNA may be treated with a restriction enzyme that cuts the DNA, and DNA fragments thus generated contain either blunt ends or overhangs known as sticky ends, and vector DNA and foreign DNA with compatible ends can then be joined by molecular ligation. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.

<span class="mw-page-title-main">Prophage</span>

A prophage is a bacteriophage genome that is integrated into the circular bacterial chromosome or exists as an extrachromosomal plasmid within the bacterial cell. Integration of prophages into the bacterial host is the characteristic step of the lysogenic cycle of temperate phages. Prophages remain latent in the genome through multiple cell divisions until activation by an external factor, such as UV light, leading to production of new phage particles that will lyse the cell and spread. As ubiquitous mobile genetic elements, prophages play important roles in bacterial genetics and evolution, such as in the acquisition of virulence factors.

<span class="mw-page-title-main">Lytic cycle</span> Cycle of viral reproduction

The lytic cycle is one of the two cycles of viral reproduction, the other being the lysogenic cycle. The lytic cycle results in the destruction of the infected cell and its membrane. Bacteriophages that only use the lytic cycle are called virulent phages.

A phagemid or phasmid is a DNA-based cloning vector, which has both bacteriophage and plasmid properties. These vectors carry, in addition to the origin of plasmid replication, an origin of replication derived from bacteriophage. Unlike commonly used plasmids, phagemid vectors differ by having the ability to be packaged into the capsid of a bacteriophage, due to their having a genetic sequence that signals for packaging. Phagemids are used in a variety of biotechnology applications; for example, they can be used in a molecular biology technique called "Phage Display".

A cosmid is a type of hybrid plasmid that contains a Lambda phage cos sequence. They are often used as a cloning vector in genetic engineering. Cosmids can be used to build genomic libraries. They were first described by Collins and Hohn in 1978. Cosmids can contain 37 to 52 kb of DNA, limits based on the normal bacteriophage packaging size. They can replicate as plasmids if they have a suitable origin of replication (ori): for example SV40 ori in mammalian cells, ColE1 ori for double-stranded DNA replication, or f1 ori for single-stranded DNA replication in prokaryotes. They frequently also contain a gene for selection such as antibiotic resistance, so that the transformed cells can be identified by plating on a medium containing the antibiotic. Those cells which did not take up the cosmid would be unable to grow.

<span class="mw-page-title-main">Lysogenic cycle</span> Process of virus reproduction

Lysogeny, or the lysogenic cycle, is one of two cycles of viral reproduction. Lysogeny is characterized by integration of the bacteriophage nucleic acid into the host bacterium's genome or formation of a circular replicon in the bacterial cytoplasm. In this condition the bacterium continues to live and reproduce normally, while the bacteriophage lies in a dormant state in the host cell. The genetic material of the bacteriophage, called a prophage, can be transmitted to daughter cells at each subsequent cell division, and later events can release it, causing proliferation of new phages via the lytic cycle. Lysogenic cycles can also occur in eukaryotes, although the method of DNA incorporation is not fully understood. For instance the AIDS viruses can either infect humans lytically, or lay dormant (lysogenic) as part of the infected cells' genome, keeping the ability to return to lysis at a later time. The rest of this article is about lysogeny in bacterial hosts.

A genomic library is a collection of overlapping DNA fragments that together make up the total genomic DNA of a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. The fragments are then inserted into the vector using DNA ligase. Next, the vector DNA can be taken up by a host organism - commonly a population of Escherichia coli or yeast - with each cell containing only one vector molecule. Using a host cell to carry the vector allows for easy amplification and retrieval of specific clones from the library for analysis.

Viral vectors are tools commonly used by molecular biologists to deliver genetic material into cells. This process can be performed inside a living organism or in cell culture. Viruses have evolved specialized molecular mechanisms to efficiently transport their genomes inside the cells they infect. Delivery of genes or other genetic material by a vector is termed transduction and the infected cells are described as transduced. Molecular biologists first harnessed this machinery in the 1970s. Paul Berg used a modified SV40 virus containing DNA from the bacteriophage λ to infect monkey kidney cells maintained in culture.

Salmonella virus P22 is a bacteriophage in the Podoviridae family that infects Salmonella typhimurium. Like many phages, it has been used in molecular biology to induce mutations in cultured bacteria and to introduce foreign genetic material. P22 has been used in generalized transduction and is an important tool for investigating Salmonella genetics.

P1 is a temperate bacteriophage that infects Escherichia coli and some other bacteria. When undergoing a lysogenic cycle the phage genome exists as a plasmid in the bacterium unlike other phages that integrate into the host DNA. P1 has an icosahedral head containing the DNA attached to a contractile tail with six tail fibers. The P1 phage has gained research interest because it can be used to transfer DNA from one bacterial cell to another in a process known as transduction. As it replicates during its lytic cycle it captures fragments of the host chromosome. If the resulting viral particles are used to infect a different host the captured DNA fragments can be integrated into the new host's genome. This method of in vivo genetic engineering was widely used for many years and is still used today, though to a lesser extent. P1 can also be used to create the P1-derived artificial chromosome cloning vector which can carry relatively large fragments of DNA. P1 encodes a site-specific recombinase, Cre, that is widely used to carry out cell-specific or time-specific DNA recombination by flanking the target DNA with loxP sites.

<span class="mw-page-title-main">Mobilome</span>

The mobilome is the entire set of mobile genetic elements in a genome. Mobilomes are found in eukaryotes, prokaryotes, and viruses. The compositions of mobilomes differ among lineages of life, with transposable elements being the major mobile elements in eukaryotes, and plasmids and prophages being the major types in prokaryotes. Virophages contribute to the viral mobilome.

A P1-derived artificial chromosome, or PAC, is a DNA construct derived from the DNA of P1 bacteriophages and Bacterial artificial chromosome. It can carry large amounts of other sequences for a variety of bioengineering purposes in bacteria. It is one type of the efficient cloning vector used to clone DNA fragments in Escherichia coli cells.

In molecular cloning, a vector is any particle used as a vehicle to artificially carry a foreign nucleic sequence – usually DNA – into another cell, where it can be replicated and/or expressed. A vector containing foreign DNA is termed recombinant DNA. The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors are an origin of replication, a multicloning site, and a selectable marker.

<span class="mw-page-title-main">Bacteriophage P2</span> Species of virus

Bacteriophage P2, scientific name Escherichia virus P2, is a temperate phage that infects E. coli. It is a tailed virus with a contractile sheath and is thus classified in the genus Peduovirus, subfamily Peduovirinae, family Myoviridae within order Caudovirales. This genus of viruses includes many P2-like phages as well as the satellite phage P4.

<span class="mw-page-title-main">Gene transfer agent</span> DNA-containing virus-like particles produced by bacteria and archaea

Gene transfer agents (GTAs) are DNA-containing virus-like particles that are produced by some bacteria and archaea and mediate horizontal gene transfer. Different GTA types have originated independently from viruses in several bacterial and archaeal lineages. These cells produce GTA particles containing short segments of the DNA present in the cell. After the particles are released from the producer cell, they can attach to related cells and inject their DNA into the cytoplasm. The DNA can then become part of the recipient cells' genome.

The CTXφ bacteriophage is a filamentous bacteriophage. It is a positive-strand DNA virus with single-stranded DNA (ssDNA).

Escherichia virus CC31, formerly known as Enterobacter virus CC31, is a dsDNA bacteriophage of the subfamily Tevenvirinae responsible for infecting the bacteria family of Enterobacteriaceae. It is one of two discovered viruses of the genus Karamvirus, diverging away from the previously discovered T4virus, as a clonal complex (CC). CC31 was first isolated from Escherichia coli B strain S/6/4 and is primarily associated with Escherichia, even though is named after Enterobacter.

References

  1. Transduction, Genetic at the U.S. National Library of Medicine Medical Subject Headings (MeSH)
  2. Jones E, Hartl DL (1998). Genetics: principles and analysis . Boston: Jones and Bartlett Publishers. ISBN   978-0-7637-0489-6.
  3. Zinder ND, Lederberg J (November 1952). "Genetic exchange in Salmonella". Journal of Bacteriology. 64 (5): 679–99. doi:10.1128/JB.64.5.679-699.1952. PMC   169409 . PMID   12999698.
  4. Snyder L, Peters JE, Henkin TM, Champness W (2013). "Lysogeny: the λ Paradigm and the Role of Lysogenic Conversion in Bacterial Pathogenesis". Molecular Genetics of Bacteria (4th ed.). Washington, DC: ASM Press. pp. 340–343. ISBN   9781555816278.
  5. Chen J.; et al. (13 October 2018). "Genome hypermobility by lateral transduction". Science. 362 (6411): 207–212. Bibcode:2018Sci...362..207C. doi: 10.1126/science.aat5867 . PMID   30309949.
  6. Denning W, Das S, Guo S, Xu J, Kappes JC, Hel Z (March 2013). "Optimization of the transductional efficiency of lentiviral vectors: effect of sera and polycations". Molecular Biotechnology. 53 (3): 308–14. doi:10.1007/s12033-012-9528-5. PMC   3456965 . PMID   22407723.