The Gateway cloning method, invented and commercialized by Invitrogen since the late 1990s, is the cloning method of the integration and excision recombination reactions that take place when bacteriophage lambda infects bacteria. This technology provides a fast and highly efficient way to transport DNA sequences into multi-vector systems for functional analysis and protein expression using Gateway att sites, and two proprietary enzyme mixes called BP Clonase and LR Clonase. In vivo, these recombination reactions are facilitated by the recombination of attachment sites from the lambda/phage chromosome (attP) and the bacteria (attB). As a result of recombination between the attP and attB sites, the phage integrates into the bacterial genome flanked by two new recombination sites (attLeft and attRight). The removal of the phage from the bacterial chromosome and the regeneration of attP and attB sites can both result from the attL and attR sites recombining under specific circumstances.
DNA sequences to be cloned are added to modified versions of these special Gateway Att sites. Two enzyme reactions take place, BP Clonase and LR Clonase. The BP Clonase occurs between the attB sites surrounding the insert and the attP sites of the donor vector. This reaction is catalyzed by the BP Clonase enzyme mixture and produces the entry clone containing the DNA of interest flanked by attL domains. As a byproduct of the reaction, the ccdB gene is cut from the donor vector. The LR Clonase occurs between the attL regions of the generated entry clone and the attR regions of the target vector and is catalyzed by the LR Clonase enzyme mix. As a result, an expression clone with DNA of interest flanked by attB regions is produced. As in the BP reaction, a DNA sequence containing the ccdB gene is cut from the target vector.
Large archives of Gateway Entry clones, containing the vast majority of human, mouse and rat ORFs (open reading frames) have been cloned from human cDNA libraries or chemically synthesized to support the research community using NIH (National Institutes of Health) funding (e.g., Mammalian Gene Collection, http://mgc.nci.nih.gov/ Archived 2015-02-25 at the Wayback Machine). The availability of these gene cassettes in a standard Gateway cloning plasmid helps researchers quickly transfer these cassettes into plasmids that facilitate the analysis of gene function. Gateway cloning does take more time for initial set-up, and is more expensive than traditional restriction enzyme and ligase-based cloning methods, but it saves time, and offers simpler and highly efficient cloning for down-stream applications.
The technology has been widely adopted by the life science research community especially for applications that require the transfer of thousands of DNA fragments into one type of plasmid (e.g., one containing a CMV promoter for protein expression in mammalian cells), or for the transfer of one DNA fragment into many different types of plasmids (e.g., for bacterial, insect and mammalian protein expression).
and 24 Related for: Gateway Technology information
The Gateway cloning method, invented and commercialized by Invitrogen since the late 1990s, is the cloning method of the integration and excision recombination...
Look up gateway in Wiktionary, the free dictionary. Gateway often refers to: A gate or portal Gateway or The Gateway may also refer to: Gateway (film)...
Gateway Motorsports Park (now known as World Wide Technology Raceway) is a motorsport racing facility in Madison, Illinois, just east of St. Louis, close...
A payment gateway is a merchant service provided by an e-commerce application service provider that authorizes credit card or direct payment processing...
The Gateway of India is an arch-monument completed in 1924 on the waterfront of Mumbai (Bombay), India. It was erected to commemorate the landing of George...
Gateway STEM High School (formerly known as Gateway Institute of Technology) is a public magnet high school in St. Louis, Missouri, United States. Gateway...
A default gateway is the node in a computer network using the Internet protocol suite that serves as the forwarding host (router) to other networks when...
Lunar Gateway, or simply Gateway, is a space station which Artemis program participants plan to assemble in an orbit near the Moon. The Gateway is intended...
The Gateway Arch is a 630-foot-tall (192 m) monument in St. Louis, Missouri, United States. Clad in stainless steel and built in the form of a weighted...
The Gateway Pundit (TGP) is an American far-right fake news website. The website is known for publishing falsehoods, hoaxes, and conspiracy theories....
of networking technologies, southbound relates to the storage used by a gateway, whereas northbound is the storage provided by the gateway). As such, one...
is a United States Air Force (USAF) airborne communications relay and gateway system carried by the unmanned EQ-4B and the manned Bombardier E-11A aircraft...
(band), an Australian post-punk/garage rock band Total Control, media gatewaytechnology created by U.S. Robotics and used by CommWorks Corporation Total Control...
once again intent on developing and experimenting with dimensional gatewaytechnology. They set up a base on one of the moons of Jupiter, with a solid detachment...
from a proposal of Yuri Gleba. It shall refer on the one hand to the GatewayTechnology, on the other hand picture the higher precision with a bridge connecting...
Border Gateway Protocol (BGP) is a standardized exterior gateway protocol designed to exchange routing and reachability information among autonomous systems...
was formerly known as GatewayTechnologies, Inc. and changed its name to Outlast Technologies, Inc. in 1997. Outlast Technologies, Inc. was founded in...
A media gateway is a translation device or service that converts media streams between disparate telecommunications technologies such as POTS, SS7, Next...
mobile gateway for open internet access to users with WAP-enabled handsets in the UK in 1998. In December 1999 Jingo took its gatewaytechnology and content...
49768 Gateway IT Park is a proposed information technology (IT) park located in the city of Kandlakoya, Medchal-Malkajgiri district, India. Gateway IT Park...
Enjoy Illinois 300 is a NASCAR Cup Series race at World Wide Technology Raceway (formerly Gateway Motorsports Park) in Madison, Illinois in the Greater St...
design and assembly Because Gateway cloning is a proprietary technology, all Gateway reactions must be carried out with the Gateway kit that is provided by...
WWT announced a naming rights sponsorship of Gateway Motorsports Park, now known as the World Wide Technology Raceway WWT also sponsors PGA golfer Graeme...