Gateway Destination Vectors

Size (kb) PCR DNA (fmol) PCR DNA (ng) Colonies/ml Transformation Correct Clones/ Total Clones Examined 

DNA from the resulting colonies is then screened for the correct cassette orientation (the ligation is bi-directional) by restriction enzymes or PCR. Once the properly oriented clone is isolated, a destination vector is born. 

For a complete list of commercially available destination vectors, please visit uniw.in-vitrogen.com. These cover a broad array of applications including protein expression in E. coli, yeast, insect, and mammalian cells, yeast two hybrid protein interaction, and gene knock-down RNAi vectors. 

Common baculoviral expression systems require the use of a bacterial transfer vector to transfer an ORF of interest into the bacuiovirus genome by homologous recombination in bacteria. This requires that the reaction be carried out in E. coli cells that have fully functional recombination machinery (this is disabled in standard 

The generation of soluble, active protein is now more important than ever with the increased emphasis on expression of proteins for structural analysis. One class of proteins that has gained importance as druggable targets are human membrane proteins. Currently, baculoviral expression is one of the best avenues for the generation of soluble membrane proteins. Therefore, easier access to baculovirus expres- 

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Many destination vectors are commercially available. However, it is also possible to construct a destination vector that is suitable for individual experiments. To constrnct snch vectors, one should insert the rr ffi-containg cassette (commercially available from Invitrogen) into the appropriate locns of a Gateway-incompatible vector. The resultant plasmids can be amplified only in the DB3.I strain, due to the toxic ccd gene (7). 

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