Novel Gateway cloning system with molecular tags in Drosophila
Gateway technology is considered to be a wonder tool in molecular biology providing faster ,easier and accurate way of cloning your gene of interest in study. This efficient technology uses att recombination sites and clonase enzyme brings in the needed recombination. To perform a gateway cloning first step will be to get the gene of interest in Entry vector to make a entry clone and later can be easily transferred into destination vectors ( with tags ) by performing LR reactions catalysed clonase enzyme. ( For more details on Gateway technology refer Davidson univ webpage and Invitrogen official webpage
Gateway technology offers many advantages over conventional cloning strategies involving restriction enzymes and few are listed below:
1) Extremely rapid with no overnight incubations
2) Easy to get your gene of interest into vector systems due to the Gateway recombination sites (attB)
3) Efficiency is almost near to 100%
4) All types of DNA fragments may be cloned, be PCR fragments, cDNA, or genomic DNA. Gateway is also available for several organisms, from mammals and insects to E. coli.
Recently Omar S. Akbari, Daniel Oliver, Katie Eyer, and Chi-Yun Pai modified this already efficient technology for expressing dosage-sensitive proteins in Drosophila melanogaster. Its no secret about the worth of tagged proteins in biology to follow the path of biomolecules in living cells.
In Drosophila UAS Gal4 system offers an excellent way to over express a protein in the desired tissue but most often than not over-expression of fusion proteins sometimes leads to the unwanted lethality or developmental defects. To overcome this problem Omar Akbari et al, developed a set of Entry/GatewayÂ® vectors for expressing fluorescent fusion proteins in Drosophila melanogaster at physiological levels.
Read the original article which published in BMC cell biology
Reference and Image Credit :
An Entry/Gateway cloning system for general expression of genes with molecular tags in Drosophila melanogaster.
Omar S Akbari , Daniel Oliver, Katie Eyer and Chi-Yun Pai
BMC Cell Biology 2009, 10:8doi:10.1186/1471-2121-10-8