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Binary Fission

JoVE 10759

Fission is the division of a single entity into two or more parts, which regenerate into separate entities that resemble the original. Organisms in the Archaea and Bacteria domains reproduce using binary fission, in which a parent cell splits into two parts that can each grow to the size of the original parent cell. This asexual method of reproduction produces cells that are all genetically identical. Though its speed varies among species, binary fission is generally rapid and can yield staggering growth. In the amount of time it takes bacterial cells to undergo binary fission, the number of cells in the bacterial culture doubles. Thus, this period is the doubling time. For example, Escherichia coli cells typically divide every 20 minutes. Bacterial growth, however, is limited by factors including nutrient and space availability. Thus, binary fission occurs at much lower rates in bacterial cultures that have encountered a growth-limiting factor (i.e., entered a stationary growth phase). In addition to organisms in the Archaea and Bacteria domains, some organelles in eukaryotic cells also reproduce via binary fission. Mitochondria, for example, divide by prokaryotic binary fission. This process requires the division of mitochondrial proteins and DNA.

 Core: Biology

Genomic DNA in Prokaryotes

JoVE 10758

The genome of most prokaryotic organisms consists of double-stranded DNA organized into one circular chromosome in a region of cytoplasm called the nucleoid. The chromosome is tightly wound, or supercoiled, for efficient storage. Prokaryotes also contain other circular pieces of DNA called plasmids. These plasmids are smaller than the chromosome and often carry genes that confer adaptive functions, such as antibiotic resistance. Although bacterial genomes are much smaller than eukaryotic genomes, they vary considerably in size and gene content. One of the smallest known bacterial genomes is that of Mycoplasma genitalium, a sexually transmitted pathogen that causes urinary and genital tract infections in humans. The M. genitalium genome is 580,076 base pairs long and consists of 559 (476 coding and 83 noncoding) genes. On the other end of the spectrum lies a particular strain of Sorangium cellulosum, a soil-dwelling bacterium. The S. cellulosum genome is enormous for a bacterium at 14,782,125 base pairs long, encoding 11,599 genes. Before the discovery of antibiotics, minor injuries could turn deadly due to the inability to stop simple bacterial infections. The discovery of penicillin in 1928 ushered in the antibiotic era, characterized by revolutionizing medical treatments and an increase in life expectancy. Howe

 Core: Biology

Serial Dilutions and Plating: Microbial Enumeration

JoVE 10507

Source: Jonathan F. Blaize1, Elizabeth Suter1, and Christopher P. Corbo1
1 Department of Biological Sciences, Wagner College, 1 Campus Road, Staten Island NY, 10301

Quantitative assessment of prokaryotes can be onerous given their abundance, propensity for exponential proliferation, species diversity within a population, and specific …


Plasmid Purification

JoVE 5062

Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under…

 Basic Methods in Cellular and Molecular Biology

Microbial and Fungal Diversity- Concept

JoVE 10601

Bacteria and fungi are two highly diverse groups of organisms that can have significant beneficial or detrimental impacts on human health. For this reason, it is important to understand and distinguish between individual species of these groups. As you will recall, biological taxonomists group organisms based on their phylogenetic relatedness. The three domains of life, Bacteria, Archaea, and…

 Lab Bio

Microbial and Fungal Diversity - Prep Student

JoVE 10638

Nutrient Agar and Bacterial Culture Plate Preparation
Add 23 g of nutrient agar to a 2,000 mL beaker containing 1 L of distilled water 4 – 5 days before the activity.
Place the beaker on a hotplate and set the hotplate to high, stirring the mixture with a stirring rod.
When the agar has dissolved into the water, use a hot pad to…

 Lab Bio

Lysogenic Cycle of Bacteriophages

JoVE 10824

In contrast to the lytic cycle, phages infecting bacteria via the lysogenic cycle do not immediately kill their host cell. Instead, they combine their genome with the host genome, allowing the bacteria to replicate the phage DNA along with the bacterial genome. The incorporated copy of the phage genome is called the prophage. Some prophages can re-activate and enter the lytic cycle. This often occurs in response to a perturbation, such as DNA damage, but can also transpire in the absence of external cues. In some cases, the genes encoded by prophages can alter the phenotype of the infected bacterium, a process known as lysogenic conversion. Some phages encode proteins or toxins called virulence factors that can facilitate bacterial infections. Through lysogenic conversion, normally non-pathogenic bacteria can become highly virulent via infection by a phage carrying virulence factors. For example, such phages are largely responsible for the pathogenicity of the bacterial species that cause botulism (Clostridium botulinum), diphtheria (Corynebacterium diphtheriae), and cholera (Vibrio cholerae). Without lysogenic conversion, these bacteria do not usually cause disease. A particularly well-studied example of lysogenic conversion is that of the Escherichia coli strain O157:H7. Several massive food recalls have stemmed

 Core: Biology


JoVE 10984

Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by a repressor protein. Altogether, the promoter, operator, structural genes, and terminator form the core of an operon. Operons are usually either inducible or repressible. Inducible operons, such as the bacterial lac operon, are normally “off” but will turn “on” in the presence of a small molecule called an inducer (e.g., allolactose). When glucose is absent, but lactose is present, allolactose binds and inactivates the lac operon repressor—allowing the operon to generate enzymes responsible for lactose metabolism. Repressible operons, such as the bacterial trp operon, are usually “on” but will turn “off” in the presence of a small molecule called a corepressor (e.g., tryptophan). When tryptophan—an essential amino acid—is abundant, tryptophan binds and activates the

 Core: Biology

The Central Dogma

JoVE 10798

The central dogma of biology states that information encoded in the DNA is transferred to messenger RNA (mRNA), which then directs the synthesis of protein. The set of instructions that enable the mRNA nucleotide sequence to be decoded into amino acids is called the genetic code. The universal nature of this genetic code has spurred advances in scientific research, agriculture, and medicine. In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is directly converted into its encoded protein. Two crucial discoveries in eukaryotic cells challenged this theory: First, protein production does not take place in the nucleus. Second, DNA is not present outside the nucleus. These findings sparked the search for an intermediary molecule that connects DNA with protein production. This intermediary molecule, found in both the nucleus and the cytoplasm, and associated with protein production, is RNA. During transcription, RNA is synthesized in the nucleus, using DNA as a template. The newly-synthesized RNA is similar in sequence to the DNA strand, except thymidine in DNA is replaced by uracil i

 Core: Biology
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