Yeast Two-Hybrid System

Yeast Two-Hybrid System is a molecular biology technique which tests the physical interactions (such as binding) between two proteins or a single protein and a DNA molecular to identify protein-protein interactions and protein-DNA interactions, respectively.

The premise behind the Yeast Two-Hybrid System is that most eukaryotic transcription factors are modular. The expression of report gene(s) is actived by the binding of transcription factor onto an upstream activating sequence (UAS). The transcription factor is split into two separate fragments, DNA-binding domain (BD, which directly binds to the UAS) and activating domain (AD, which recruts the transcriptional machinery).

Yeast Two-Hybrid System exploit the modular nature of the yeast Gal4 transcription factor to detect possible molecular interactions. In this assay, the DNA-binding domain fragment of Gal4 is fused to a protein of interest called the bait, while activation domain fragment is fused to number of potential binding partners called the prey. If the bait and prey interact, an active Gal4 transcription factor is reconstituted to activate the expression of reporter genes under the control of the GAL promoter. If the bait and prey do not interact, the two parts of the transcription factor remain separate and activation of gene expression doesn’t occur.

Fig 1. Principle of Yest Two-Hybrid System

• Yeast one-hybrid: Idenfication of protein-DNA interactions;
• Yeast three-hybrid: Identification of protein-RNA interactions;
• Split ubiquitin yeast two-hybrid: Identification of protein-protein interactions between non-soluble proteins, such as membrane proteins;
• Yeast one-two-hybrid: Identification of simultaneous protein-DNA and protein-protein interactions.

• Determination of sequences crucial for interaction: Determine the importance of some amino acid residues in maintaining the interaction by changing them by mutating the corresponding DNA base-pairs in the plasmids used.
• Determination of protein function: Determine the possible functions of new proteins by deteting the interaction partners of unknown proteins.
• Drug discovery: Screen the functional site of new drugs and the effection of the drug to the interaction of target protein.
• Study of interaction between antigen and antibody in vivo
• Establishment of genomic protein linkage map

Yeast Two-Hybrid System is used to identify protein interactions in a living yeast cell, it offers a number of advantages.

• Simple: Eliminate the need for purification of protein.
• Low-tech: Can be carried out in any lab without sophisticated equipment.
• Scalable: Enable screening interactions among many proteins.
• Fast: Ablt to identify the interaction of many proteins with thousands of potentially interacting proteins in a relatively short time by using robots.
• Sensitive: Able to detect the weak or short-time interactions between proteins thanks to the cumulative effect of gene expression products.
• Economic: Provide similar data generated by MS-based coaffinity purification which requires expensive large equipment

Pipettes

A pipette is a laboratory tool commonly used in chemistry, biology and medicine to transport a measured volume of liquid, often as a media dispenser. Pipettes come in several designs for various purposes with differing levels of accuracy and precision, from single piece glass pipettes to more complex adjustable or electronic pipettes.

Laminar Flow Hoods

Laminar Flow Hoods are mostly enclosed systems that use filters and directrional air flow to provide a contaminant-free work area for applications requiring a sterile work space, such as tissue culture, culture media preparation, and so on.

Ice Maker

An ice maker is a machine for making ice. Some types of laboratory experiments and reactions require an ice bath in one or more of its steps. Lab ice makers are a convenient way to ensure ice is there when it’s needed. An ice maker is very useful in microbiology laboratories and for PCR (polymerase chain reaction).

Vortexer

Vortexer is a simple device used commonly in laboratories to mix small vials of liquid. Vortexers are quite commonplace in bioscience laboratories. In cell culture and microbiology laboratories they may be used to suspend cells. In a biochemical or analytical laboratory they may be used to mix the reagents of an assay or to mix an experimental sample and a dilutant.

Benchtop Centrifuges

Centrifuges separate or concentrate substances suspended in a liquid medium by density. Space-saving fixed- and variable-speed benchtop or tabletop centrifuges are used for applications including tissue culture, protein work, DNA/RNA research, and cell harvesting.

Thermal Cyclers

Thermal Cycler is a laboratory apparatus that amplify segments of DNA using PCR. A thermal cycler is also commonly called a DNA Amplifier, PCR Machine, or Thermocycler. The cycle normally lowers or raises the block’s temperature in preprogrammed discrete steps. A thermal cycler is important for a laboratory dealing in molecular biology and gene cloning.

Laboratory Rockers and Shakers

A rocker or shaker is likely to be found on the bench top of most laboratories whether small, large, academic, clinical or commercial. These important tools are used for a variety of applications including cell culture, DNA extraction, low foaming agitation, mixing reagents, and staining gels and blots.

Laboratory Incubators

Laboratory Incubators provide a controlled, contaminant-free environment for safe, reliable work with cell and tissue cultures by regulating conditions such as temperature, humidity, and CO2. Microbiological incubators are used for the growth and storage of bacterial cultures.

Electrophoresis

Electrophoresis is a laboratory technique used to separate DNA, RNA or protein molecules based on their size and electrical charge.

STEMart offers complete instruments and equipments used in Yeast Two-Hybrid System to facilitate your research. If you want to learn more detail about the related devices, or would like to consult with the experts at STEMart, please feel free to contact us.