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A marker gene
is a gene
A gene is a molecular unit of heredity of a living organism. It is a name given to some stretches of DNA and RNA that code for a type of protein or for an RNA chain that has a function in the organism. Living beings depend on genes, as they specify all proteins and functional RNA chains...
used in molecular biology
Molecular biology is the branch of biology that deals with the molecular basis of biological activity. This field overlaps with other areas of biology and chemistry, particularly genetics and biochemistry...
to determine if a nucleic acid sequence has been successfully inserted into an organism's DNA
Deoxyribonucleic acid is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms . The DNA segments that carry this genetic information are called genes, but other DNA sequences have structural purposes, or are involved in...
. There are two types of marker genes: a selectable marker
and a marker for screening
A selectable marker protects the organism from a selective agent
that would normally kill it or prevent its growth. In most applications, only one in a several billion cells will take up DNA. Rather than checking every single cell, scientists use a selective agent to kill all cells that do not contain the foreign DNA, leaving only the desired ones.
An antibacterial is a compound or substance that kills or slows down the growth of bacteria.The term is often used synonymously with the term antibiotic; today, however, with increased knowledge of the causative agents of various infectious diseases, antibiotic has come to denote a broader range of...
s are the most common selective agents. In bacteria, antibiotics are used almost exclusively. In plants, antibiotics that kill the chloroplast
Chloroplasts are organelles found in plant cells and other eukaryotic organisms that conduct photosynthesis. Chloroplasts capture light energy to conserve free energy in the form of ATP and reduce NADP to NADPH through a complex set of processes called photosynthesis.Chloroplasts are green...
are often used as well, although tolerance to salts and growth-inhibiting hormones is becoming more popular. In mammals, resistance to antibiotics that would kill the mitochondria is used as a selectable marker.
Marker for screening
A marker for screening will make cells containing the gene look different. There are three types of screening commonly used:
- Green fluorescent protein
The green fluorescent protein is a protein composed of 238 amino acid residues that exhibits bright green fluorescence when exposed to blue light. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the...
makes cells glow green under UV light. A specialized microscope is required to see individual cells. Yellow and red versions are also available, so scientists can look at multiple genes at once. It is commonly used to measure gene expression
Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product. These products are often proteins, but in non-protein coding genes such as ribosomal RNA , transfer RNA or small nuclear RNA genes, the product is a functional RNA...
- GUS assay (using β-glucuronidase) is an excellent method for detecting a single cell by staining it blue without using any complicated equipment. The drawback is that the cells are killed in the process. It is particularly common in plant science.
- Blue/white screening is used in both bacteria and eukaryotic cells. The bacterial lacZ gene encodes a beta-galactosidase
β-galactosidase, also called beta-gal or β-gal, is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides. Substrates of different β-galactosidases include ganglioside GM1, lactosylceramides, lactose, and various glycoproteins...
enzyme. When media containing certain galactosides (e.g. X-gal
X-gal is an organic compound consisting of galactose linked to a substituted indole. The compound was synthesized by Jerome Horwitz and collaborators in Detroit, MI, in 1964. The formal chemical name is often shortened to less accurate but also less cumbersome phrases such as bromochloroindoxyl...
) is added, cells expressing the gene convert the X-gal to a blue product and can be seen with the naked eye.