Microbial Genetics
Microbial genetics
Microbial genetics is a branch of genetics concerned with the transmission of hereditary characters in microorganisms . Within the usual definition, microorganisms include prokaryotes like bacteria , unicellular or mycelial eukaryotes e.g., yeasts and other fungi , and viruses , notably bacterial viruses (bacteriophages). Microbial genetics has played a unique role in developing the fields of molecular and cell biology and also has found applications in medicine, agriculture, and the food and pharmaceutical industries.
Because of their relative simplicity, microbes are ideally suited for combined biochemical and genetic studies, and have been successful in providing information on the genetic code and the regulation of gene activity. The operon model formulated by French biologists François Jacob (1920– ) and Jacques Monod (1910–1976) in 1961, is one well known example. Based on studies on the induction of enzymes of lactose catabolism in the bacterium Escherichia coli, the operon has provided the groundwork for studies on gene expression and regulation, even up to the present day. The many applications of microbial genetics in medicine and the pharmaceutical industry emerge from the fact that microbes are both the causes of disease and the producers of antibiotics . Genetic studies have been used to understand variation in pathogenic microbes and also to increase the yield of antibiotics from other microbes.
Hereditary processes in microorganisms are analogous to those in multicellular organisms. In both prokaryotic and eukaryotic microbes, the genetic material is DNA ; the only known exceptions to this rule are the RNA viruses. Mutations , heritable changes in the DNA, occur spontaneously and the rate of mutation can be increased by mutagenic agents. In practice, the susceptibility of bacteria to mutagenic agents has been used to identify potentially hazardous chemicals in the environment. For example, the Ames test was developed to evaluate the mutagenicity of a chemical in the following way. Plates containing a medium lacking in, for example, the nutrient histidine are inolculated with a histidine requiring strain of the bacterium Salmonella typhimurium. Thus, only cells that revert back to the wild type can grow on the medium. If plates are exposed to a mutagenic agent, the increase in the number of mutants compared with unexposed plates can be observed and a large number of revertants would indicate a strong mutagenic agent. For such studies, microorganisms offer the advantage that they have short mean generation times, are easily cultured in a small space under controlled conditions and have a relatively uncomplicated structure.
Microorganisms, and particularly bacteria, were generally ignored by the early geneticists because of their small in size and apparent lack of easily identifiable variable traits. Therefore, a method of identifying variation and mutation in microbes was fundamental for progress in microbial genetics. As many of the mutations manifest themselves as metabolic abnormalities, methods were developed by which microbial mutants could be detected by selecting or testing for altered phenotypes. Positive selection is defined as the detection of mutant cells and the rejection of unmutated cells. An example of this is the selection of penicillin resistant mutants, achieved by growing organisms in media containing penicillin such that only resistant colonies grow. In contrast, negative selection detects cells that cannot perform a certain function and is used to select mutants that require one or more extra growth factors. Replica plating is used for negative selection and involves two identical prints of colony distributions being made on plates with and without the required nutrients. Those microbes that do not grow on the plate lacking the nutrient can then be selected from the identical plate, which does contain the nutrient.
The first attempts to use microbes for genetic studies were made in the USA shortly before World War II, when George W. Beadle (1903–1989) and Edward L. Tatum (1909–1975) employed the fungus, Neurospora, to investigate the genetics of tryptophan metabolism and nicotinic acid synthesis. This work led to the development of the "one gene one enzyme" hypothesis. Work with bacterial genetics, however, was not really begun until the late 1940s. For a long time, bacteria were thought to lack sexual reproduction, which was believed to be necessary for mixing genes from different individual organisms—a process fundamental for useful genetic studies. However, in 1947, Joshua Lederberg (1925– ) working with Edward Tatum demonstrated the exchange of genetic factors in the bacterium, Escherichia coli. This process of DNA transfer was termed conjugation and requires cell-to-cell contact between two bacteria. It is controlled by genes carried by plasmids , such as the fertility (F) factor, and typically involves the transfer of the plasmid from donor torecipient cell. Other genetic elements, however, including the donor cell chromosome, can sometimes also be mobilized and transferred. Transfer to the host chromosome is rarely complete, but can be used to map the order of genes on a bacterial genome.
Other means by which foreign genes can enter a bacterial cell include transformation , transfection, and transduction . Of the three processes, transformation is probably the most significant. Evidence of transformation in bacteria was first obtained by the British scientist, Fred Griffith (1881–1941) in the late 1920s working with Streptococcus pneumoniae and the process was later explained in the 1930s by Oswald Avery (1877–1955) and his associates at the Rockefeller Institute in New York. It was discovered that certain bacteria exhibit competence, a state in which cells are able to take up free DNA released by other bacteria. This is the process known as transformation, however, relatively few microorganisms can be naturally transformed. Certain laboratory procedures were later developed that make it possible to introduce DNA into bacteria, for example electroporation, which modifies the bacterial membrane by treatment with an electric field to facilitate DNA uptake. The latter two processes, transfection and transduction, involve the participation of viruses for nucleic acid transfer. Transfection occurs when bacteria are transformed with DNA extracted from a bacterial virus rather than from another bacterium. Transduction involves the transfer of host genes from one bacterium to another by means of viruses. In generalized transduction, defective virus particles randomly incorporate fragments of the cell DNA; virtually any gene of the donor can be transferred, although the efficiency is low. In specialized transduction, the DNA of a temperate virus excises incorrectly and brings adjacent host genes along with it. Only genes close to the integration point of the virus are transduced, and the efficiency may be high.
After the discovery of DNA transfer in bacteria, bacteria became objects of great interest to geneticists because their rate of reproduction and mutation is higher than in larger organisms; i.e., a mutation occurs in a gene about one time in 10,000,000 gene duplications, and one bacterium may produce 10,000,000,000 offspring in 48 hours. Conjugation, transformation, and transduction have been important methods for mapping the genes on the chromosomes of bacteria. These techniques, coupled with restriction enzyme analysis, cloning DNA sequencing, have allowed for the detailed studies of the bacterial chromosome. Although there are few rules governing gene location, the genes encoding enzymes for many biochemical pathways are often found tightly linked in operons in prokaryotes. Large scale sequencing projects revealed the complete DNA sequence of the genomes of several prokaryotes, even before eukaryotic genomes were considered.
See also Bacterial growth and division; Bacteriophage and bacteriophage typing; Cell cycle (eukaryotic), genetic regulation of; Cell cycle (prokaryotic), genetic regulation of; Fungal genetics; Mutations and mutagenesis; Viral genetics; Viral vectors in gene therapy
Microbial Genetics
Microbial Genetics
Microbial genetics is a branch of genetics concerned with the transmission of hereditary characters in microorganisms. Within the usual definition, microorganisms include prokaryotes like bacteria, unicellular or mycelial eukaryotes e.g., yeasts and other fungi, and viruses, notably bacterial viruses (bacteriophages). Microbial genetics has played a unique role in developing the fields of molecular and cell biology and also has found applications in medicine, agriculture, and the food and pharmaceutical industries.
Because of their relative simplicity, microbes are ideally suited for combined biochemical and genetic studies, and have been successful in providing information on the genetic code and the regulation of gene activity. The operon model formulated by French biologists François Jacob (1920–) and Jacques Monod (1910–1976) in 1961, is one well known example. Based on studies on the induction of enzymes of lactose catabolism in the bacterium Escherichia coli, the operon has provided the groundwork for studies on gene expression and regulation, even up to the present day. The many applications of microbial genetics in medicine and the pharmaceutical industry emerge from the fact that microbes are both the causes of disease and the producers of antibiotics. Genetic studies have been used to understand variation in pathogenic microbes and also to increase the yield of antibiotics from other microbes.
Hereditary processes in microorganisms are analogous to those in multicellular organisms. In both prokaryotic and eukaryotic microbes, the genetic material is DNA; the only known exceptions to this rule are the RNA viruses. Mutations, heritable changes in the DNA, occur spontaneously and the rate of mutation can be increased by mutagenic agents. In practice, the susceptibility of bacteria to mutagenic agents has been used to identify potentially hazardous chemicals in the environment. For example, the Ames test was developed to evaluate the mutagenicity of a chemical in the following way. Plates containing a medium lacking in, for example, the nutrient histidine are inolculated with a histidine requiring strain of the bacterium Salmonella typhimurium. Thus, only cells that revert back to the wild type can grow on the medium. If plates are exposed to a mutagenic agent, the increase in the number of mutants compared with unexposed plates can be observed and a large number of revertants would indicate a strong mutagenic agent. For such studies, microorganisms offer the advantage that they have short mean generation times, are easily cultured in a small space under controlled conditions and have a relatively uncomplicated structure.
Microorganisms, and particularly bacteria, were generally ignored by the early geneticists because of their small in size and apparent lack of easily identifiable variable traits. Therefore, a method of identifying variation and mutation in microbes was fundamental for progress in microbial genetics. As many of the mutations manifest themselves as metabolic abnormalities, methods were developed by which microbial mutants could be detected by selecting or testing for altered phenotypes. Positive selection is defined as the detection of mutant cells and the rejection of unmutated cells. An example of this is the selection of penicillin resistant mutants, achieved by growing organisms in media containing penicillin such that only resistant colonies grow. In contrast, negative selection detects cells that cannot perform a certain function and is used to select mutants that require one or more extra growth factors. Replica plating is used for negative selection and involves two identical prints of colony distributions being made on plates with and without the required nutrients. Those microbes that do not grow on the plate lacking the nutrient can then be selected from the identical plate, which does contain the nutrient.
The first attempts to use microbes for genetic studies were made in the United States shortly before World War II, when George W. Beadle (1903–1989) and Edward L. Tatum (1909–1975) employed the fungus, Neurospora, to investigate the genetics of tryptophan metabolism and nicotinic acid synthesis. This work led to the development of the “one-gene one-enzyme” hypothesis. Work with bacterial genetics, however, was not really begun until the late 1940s. For a long time, bacteria were thought to lack sexual reproduction, which was believed to be necessary for mixing genes from different individual organisms—a process fundamental for useful genetic studies. However, in 1947, Joshua Lederberg (1925–) working with Edward Tatum demonstrated the exchange of genetic factors in the bacterium, Escherichia coli. This process of DNA transfer was termed conjugation and requires cell-to-cell contact between two bacteria. It is controlled by genes carried by plasmids, such as the fertility (F) factor, and typically involves the transfer of the plasmid from donor to recipient cell. Other genetic elements, however, including the donor cell chromosome, can sometimes also be mobilized and transferred. Transfer to the host chromosome is rarely complete, but can be used to map the order of genes on a bacterial genome.
Other means by which foreign genes can enter a bacterial cell include transformation, transfection, and transduction. Of the three processes, transformation is probably the most significant. Evidence of transformation in bacteria was first obtained by the British scientist, Fred Griffith (1881–1941) in the late 1920s working with Streptococcus pneumoniae and the process was later explained in the 1930s by Oswald Avery (1877–1955) and his associates at the Rockefeller Institute in New York. It was discovered that certain bacteria exhibit competence, a state in which cells are able to take up free DNA released by other bacteria. This is the process known as transformation, however, relatively few microorganisms can be naturally transformed.
Certain laboratory procedures were later developed that make it possible to introduce DNA into bacteria, for example electroporation, which modifies the bacterial membrane by treatment with an electric field to facilitate DNA uptake. The latter two processes, transfection and transduction, involve the participation of viruses for nucleic acid transfer. Transfection occurs when bacteria are transformed with DNA extracted from a bacterial virus rather than from another bacterium. Transduction involves the transfer of host genes from one bacterium to another by means of viruses. In generalized transduction, defective virus particles randomly incorporate fragments of the cell DNA; virtually any gene of the donor can be transferred, although the efficiency is low. In specialized transduction, the DNA of a temperate virus excises incorrectly and brings adjacent host genes along with it. Only genes close to the integration point of the virus are transduced, and the efficiency may be high.
After the discovery of DNA transfer in bacteria, bacteria became objects of great interest to geneticists because their rate of reproduction and mutation is higher than in larger organisms; i.e., a mutation occurs in a gene about one time in 10,000,000 gene duplications, and one bacterium may produce 10,000,000,000 offspring in 48 hours. Conjugation, transformation, and transduction have been important methods for mapping the genes on the chromosomes of bacteria. These techniques, coupled with restriction enzyme analysis, cloning DNA sequencing, have allowed for the detailed studies of the bacterial chromosome. Although there are few rules governing gene location, the genes encoding enzymes for many biochemical pathways are often found tightly linked in operons in prokaryotes. Large scale sequencing projects revealed the complete DNA sequence of the genomes of several prokaryotes, even before eukaryotic genomes were considered.
Resources
BOOKS
Breeze, Roger, Bruce Budowle, and Steven Schutzer. Microbial Forensics. New York: Academic Press, 2005.
Dale, Jeremy W. and Simon F. Park. Molecular Genetics of Bacteria. New York: John Wiley & Sons, 2004.
Snyder, Larry and Wendy Champness. Molecular Genetics of Bacteria. Washington, DC: ASM Press, 2002.
Judyth Sassoon
Microbial Genetics
Microbial genetics
Microorganisms include prokaryotes like bacteria , unicellular or mycelial eukaryotes e.g., yeasts and other fungi , and viruses, notably bacterial viruses (bacteriophages). Microbial genetics is concerned with the transmission of hereditary characters in microorganisms. Microbial genetics has played a unique role in developing the fields of molecular and cell biology and also has found applications in medicine, agriculture, and the food and pharmaceutical industries.
Hereditary processes in microorganisms are analogous to those in multicellular organisms. In both prokaryotic and eukaryotic microbes, the genetic material is DNA; the only known exceptions to this rule are the RNA viruses. Mutations, heritable changes in the DNA, occur spontaneously and the rate of mutation can be increased by mutagenic agents. In practice, the susceptibility of bacteria to mutagenic agents has been used to identify potentially hazardous chemicals in the environment. For example, the Ames test was developed to evaluate the mutagenicity of a chemical in the following way. Plates containing a medium lacking in, for example, the nutrient histidine are inolculated with a histidine requiring strain of the bacterium Salmonella typhimurium. Thus, only cells that revert back to the wild type can grow on the medium. If plates are exposed to a mutagenic agent, the increase in the number of mutants compared with unexposed plates can be observed and a large number of revertants would indicate a strong mutagenic agent. For such studies, microorganisms offer the advantage that they have short mean generation times, are easily cultured in a small space under controlled conditions and have a relatively uncomplicated structure.
Microbes are ideally suited for combined biochemical and genetic studies, and have been successful in providing information on the genetic code and the regulation of gene activity. The operon model formulated by French biologists François Jacob (1920–) and Jacques Monod (1910–76) in 1961, is one well known example. Based on studies on the induction of enzymes of lactose catabolism in the bacterium Escherichia coli, the operon has provided the groundwork for studies on gene expression and regulation, even up to the present day. The many applications of microbial genetics in medicine and the pharmaceutical industry emerge from the fact that microbes are both the causes of disease and the producers of antibiotics . Genetic studies have been used to understand variation in pathogenic microbes and also to increase the yield of antibiotics from other microbes.
Microorganisms, and particularly bacteria, were generally ignored by the early geneticists because of their small in size and apparent lack of easily identifiable variable traits. Therefore, a method of identifying variation and mutation in microbes was fundamental for progress in microbial genetics. As many of the mutations manifest themselves as metabolic abnormalities, methods were developed by which microbial mutants could be detected by selecting or testing for altered phenotypes. Positive selection is defined as the detection of mutant cells and the rejection of unmutated cells. An example of this is the selection of penicillin resistant mutants, achieved by growing organisms in media containing penicillin such that only resistant colonies grow. In contrast, negative selection detects cells that cannot perform a certain function and is used to select mutants that require one or more extra growth factors. Replica plating is used for negative selection and involves two identical prints of colony distributions being made on plates with and without the required nutrients . Those microbes that do not grow on the plate lacking the nutrient can then be selected from the identical plate, which does contain the nutrient.
The first attempts to use microbes for genetic studies were made in the USA shortly before World War II, when George W. Beadle (1903–1989) and Edward L. Tatum (1909–1975) employed the fungus, Neurospora, to investigate the genetics of tryptophan metabolism and nicotinic acid synthesis. This work led to the development of the "one gene one enzyme" hypothesis. Work with bacterial genetics, however, was not really begun until the late 1940s. For a long time, bacteria were thought to lack sexual reproduction , which was believed to be necessary for mixing genes from different individual organisms—a process fundamental for useful genetic studies. However, in 1947, Joshua Lederberg (1925–) working with Edward Tatum demonstrated the exchange of genetic factors in the bacterium, Escherichia coli. This process of DNA transfer was termed conjugation and requires cell-to-cell contact between two bacteria. It is controlled by genes carried by plasmids, such as the fertility (F) factor, and typically involves the transfer of the plasmid from donor torecipient cell. Other genetic elements, however, including the donor cell chromosome , can sometimes also be mobilized and transferred. Transfer to the host chromosome is rarely complete, but can be used to map the order of genes on a bacterial genome .
Other means by which foreign genes can enter a bacterial cell include transformation, transfection, and transduction. Of the three processes, transformation is probably the most significant. Evidence of transformation in bacteria was first obtained by the British scientist, Fred Griffith (1881–1941) in the late 1920s working with Streptococcus pneumoniae and the process was later explained in the 1930s by Oswald Avery (1877–1955) and his associates at the Rockefeller Institute in New York. It was discovered that certain bacteria exhibit competence, a state in which cells are able to take up free DNA released by other bacteria. This is the process known as transformation, however, relatively few microorganisms can be naturally transformed. Certain laboratory procedures were later developed that make it possible to introduce DNA into bacteria, for example electroporation, which modifies the bacterial membrane by treatment with an electric field to facilitate DNA uptake. The latter two processes, transfection and transduction, involve the participation of viruses for nucleic acid transfer. Transfection occurs when bacteria are transformed with DNA extracted from a bacterial virus rather than from another bacterium. Transduction involves the transfer of host genes from one bacterium to another by means of viruses. In generalized transduction, defective virus particles randomly incorporate fragments of the cell DNA; virtually any gene of the donor can be transferred, although the efficiency is low. In specialized transduction, the DNA of a temperate virus excises incorrectly and brings adjacent host genes along with it. Only genes close to the integration point of the virus are transduced, and the efficiency may be high.
After the discovery of DNA transfer in bacteria, bacteria became objects of great interest to geneticists because their rate of reproduction and mutation is higher than in larger organisms; i.e., a mutation occurs in a gene about one time in 10,000,000 gene duplications, and one bacterium may produce 10,000,000,000 offspring in 48 hours. Conjugation, transformation, and transduction have been important methods for mapping the genes on the chromosomes of bacteria. These techniques, coupled with restriction enzyme analysis, cloning DNA sequencing, have allowed for the detailed studies of the bacterial chromosome. Although there are few rules governing gene location, the genes encoding enzymes for many biochemical pathways are often found tightly linked in operons in prokaryotes. Large scale sequencing projects revealed the complete DNA sequence of the genomes of several prokaryotes, even before eukaryotic genomes were considered.
See also Viral genetics.
Resources
books
Flint, S.J., L.W. Enquist, R.M. Krug, et al. Principles of Virology: Molecular Biology, Pathogenesis, and Control. Washington, DC: American Society for Microbiology Press, 1999.
Lodish, H., et. al. Molecular Cell Biology, 4th ed. New York: W. H. Freeman & Co., 2000.
Stahl, F.W. We Can Sleep Later: Alfred D. Hershey and the Origins of Molecular Biology Cold Spring Harbor, NY: Cold Spring Harbor Press, 2000.
Summers, W.C. Felix d'Herelle and the Origins of Molecular Biology. New Haven: Yale University Press, 2000.
periodicals
Fraser, C.M., J. Eisen, R.D. Fleischmann, K.A. Ketchum, and S. Peterson. "Comparative Genomics and Understanding of Microbial Biology." Emerging Infectious Diseases. 6, no.5 (September-October 2000).
other
City College of San Francisco. "Requirements for Microbial Growth." 2000 [cited February 2, 2003]. <http://www.ccsf.org/Departments/Biology/growth.htm>.
Dakota Wesleyan University. "Microbial Taxonomy." 2001 [cited February 9, 2003]. <http://www.dwu.edu/biology/Mullican/micrtaxon.htm>.
Judyth Sassoon