P[acman] BAC libraries in Drosophila : A reserch tool
Hugo bellen’s lab generated Drosophila melanogaster bacterial artificial chromosome libraries with 21-kilobase and 83-kilobase inserts in the P[acman] system. P[acman] was developed by Koen Venken in Bellen’s laboratory,which allows scientists to study large chunks of DNA in living flies.Cloning huge fragments of DNA is not possible in classical P-elements and this has been over come in P[acman] that allows modification of cloned fragments by recombineering and germline transformation of genomic DNA fragments up to 133 kilobases.
The vector – officially P/phiC31 artificial chromosome for manipulation – combines different technologies: a specially designed bacterial artificial chromosome (BAC) that allows maintenance of large pieces of DNA in bacteria, recombineering that allows the manipulation of large pieces of DNA in bacteria, and the ability to insert the genomic DNA into the genome of the fly at a specific site using phiC31-mediated transgenesis.
Authors has modified the P[acman] vector to create genomic libraries, so that now a researcher can choose a gene and find the corresponding clones in the library that cover that gene. Their collaborators at Lawrence Berkeley National Laboratory, Drs. Roger Hoskins and Joseph Carlson, played a key role in the design, construction, and annotation of the libraries. Another huge advantage is that one can tag the protein as well – If you don’t know where the gene is expressed, you can tag it, put it back and locate where it is expressed. In addition can insert a single copy of a gene and rescue a mutation, or do a structure/function analysis of the gene.
This work has been published recent issue of Nature methods ,which also have some other interesting articles related to research tools.
The library is available at http://pacmanfly.org/.
Reference :
Venken, KJT, Carlson JW, Schulze KL, Pan H, He Y, Spokony R, Wan KH, Koriabine M, de Jong PJ, White KP, Bellen HJ, Hoskins RA (2009) Versatile P[acman] BAC Libraries for transgenesis studies in Drosophila melanogaster.
doi:10.1038/nmeth.1331, Nature Methods
Other articles from same issue :
High-throughput ethomics in large groups of Drosophila – pp451 – 457
Kristin Branson, Alice A Robie, John Bender, Pietro Perona & Michael H Dickinson
doi:10.1038/nmeth.1328
A toolkit for high-throughput, cross-species gene engineering in Drosophila – pp435 – 437
Radoslaw K Ejsmont, Mihail Sarov, Sylke Winkler, Kamil A Lipinski & Pavel Tomancak
doi:10.1038/nmeth.1334
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