BDGP EST and DGC Collections
The BDGP EST Collection comprises cDNAs from a number of different libraries created by BDGP (Rubin et al., 2000; Stapleton et al., 2002).
We have received these clones "as is" from the BDGP.
Clones marked Gold are full length cDNAs and were sequenced by BDGP. Where available, we offer that sequence for download.
Other EST clones have not been fully characterized and many may exist for a single gene. These clones may or may not be full length or redundant in sequence.
If your clone of interest is in the EST Collection, but marked "unavailable", it is likely a clone that the DGRC did not receive and according to BDGP, the plate no longer exists. To our knowledge, there are no other sources for these ESTs.
BDGP ORF Collection
This collection contains high quality ORF proteomic clones from BDGP in the BD Clonetech In-Fusion system. The ORFs originate from clones in the Gold Collection.
Two clones of each ORF exist, one contains the native stop codon and is referred to as ExpressR-NativeStop(NS) clone and one lacks the stop codon and is referred to as theExpressR-Open(O) clone. The NS and O forms of the clones allow one to create amino-terminal or carboxy-terminal fusion proteins, respectively. For more information on how this collection was made, visit an archive of the BDGP Universal Proteomics Resources page.
These expression clones are cloned into pDNR-Dual from Clonetech.
BDGP Tagged ORF Collection
This is a collection of expression-ready ORFs that have been tagged.
There are seven categories of tagged ORFS:
- UFO - UAS promoter with Flag-HA tag C-terminal fusion
- MXO - metallothionein promoter with TAP tag C-terminal fusion
- FMO - metallothionein promoter with Flag-HA tag C-terminal fusion
- UGS - UAS promoter with GFP tag N-terminal fusion
- URS - UAS promoter with mCherry tag N-terminal fusion
- UGO - UAS promoter with GFP tag C-terminal fusion
- URO - UAS promoter with mCherry tag C-terminal fusion
For more information on how this collection was made, visit an archive of the BDGP Universal Proteomics Resources page.
Extra information from BDGP on the FMO clones is available in the FMO clone FAQ section.
BDGP iPCR Collection
The BDGP iPCR Collection contains cDNA clones generated by BDGP using a screening method called SLIP (Self-Ligation of Inverse PCR Products). This method is utilized to recover full-length cDNAs for relatively rare and alternatively spliced transcripts of interest. This collection contains 981 genes not previously represented in the EST or DGC collections (Hoskins et al., 2005).
Clones marked Gold are full length cDNAs and were sequenced by BDGP. Where available, we offer that sequence for download.
Testis cDNA Collection
This collection is comprised of cDNAs from a library generated from testis, vas deferens, and portions of the ejaculatory apparatus and accessory glands (Andrews et. al., 2000).
Search Tips: Clone ID can be retrieved from GenBank (search GenBank by gene name or CG number). All testes cDNA clones from this collection begin with the letters "bs" and are followed by two numbers, one letter, two numbers.
CuraGen Yeast 2-Hybrid Collection
CuraGen Corporation, in collaboration with researchers at the Johns Hopkins University, Wayne State University School of Medicine and Yale University School of Medicine, completed the first comprehensive protein interaction map for a multicellular organism, Drosophila melanogaster, and published the results in the online edition and the December 5, 2003 print edition of Science.
The research used the yeast two-hybrid protocol in a high-throughput mode to capture a significant fraction of the organism's protein-protein interaction.
CuraGen Materials available through the DGRC
The following CuraGen materials are available through the DGRC:
- 10,787 AD clones in CuraGen yeast line N106r
- 10,623 BD clones in CuraGen yeast lines YULH
Both the AD and BD clones have been QC'ed by sizing and sequencing in the 5' direction.
The vectors used to make these clones were pBD-Gal4 CAM ("BD" from Stratagene) and pAct2 ("AD" from BD Biosciences).
The yeast 2-hybrid clones were generated using the primers designed based on release 1 and 2. The yeast 2-hybrid Primer File is available to download and should be used as reference only. It is our understanding that CuraGen only kept track of the gene identity by sequencing once the clones were generated; not based on the gene intended to be amplified by the primers listed.
ECD Collection
The extracellular domain (ECD) collection consists of a set of 202 extracellular domains of Drosophila proteins composed of the immunoglobulin superfamily (IgSF), fibronectin type III (FnIII), and leucine-rich repeat (LRR) families. They were each individually cloned into two complementary bait and prey vectors - to create a library of 404 clones - that permits expression and secretion from Drosophila cell lines. See Ozkan et al., 2003 for more information.
FlyBi ORFeome Collection
The FlyBi ORFeome Collection contains 10,271 open reading frames (ORFs) from D. melanogaster, cloned in the pDONR223 Gateway vector. These represent 2/3 of the proteome. Visit FlyBi at HMS for more information.
Human cDNA Collection
The Human cDNA Collection contains human orthologs in pUAS vectors for Drosophila transgenesis and cell culture. A few of the cDNAs are also available as Gateway entry clones. The collections have been transferred to DGRC by Drs. Travis Johnson, Hugo Bellen, Michael O'Connor, Stuart Newfeld and BDGP unpublished).
Tagged Transcription Factor BAC Collection
The Tagged Transcription Factor BAC Collection contains 650 clones in attB-BAC for Drosophila transgenics (Dr. Kevin White, unpublished). These are BAC clones containing Drosophila transcription factors in which recombineering was utilized to add an epitope tag to the carboxy end of the open reading frame. The transcription factor is under the control of its endogenous promoter sequences and the BACs contain an attB site for high efficiency integration into the Drosophila genome.
Dr. Kevin White has provided a spreadsheet with information on each clone.
Weizmann gRNA Collection
This collection comes from the Weizmann Knock Out project (used to create the "WKO lines") as described in Meltzer et al., 2019. It contains ~850 plasmids with gRNA sequences cloned into pCFD4 or pCFD5. Selection of genes for this gRNA library were made based on expression data of developing MB-gamma neurons.
Phosphoinositide signaling gRNA Library
Phosphoinositide signaling gRNA library was created by the Padinjat Lab (Trivedi et al., 2020). The library contains two sgRNAs cloned into either pBFv6.2 and pBFv6.2B vectors targeting each of the 103 Phosphoinositide signaling genes. For every gene, one sgRNA targets the first coding exon (sg1, cloned into pBFv6.2) and another targets the last coding exon (sg2, cloned into pBFv6.2B). For every gene targeted in the collection, dual gRNA plasmids are also available, that carry both the sgRNA's (sg1 and sg2) in a single plasmid (pBFv6.2B). The gRNA library has been validated for the ability to generate deletions in S2R+ cells and flies.
Clones
The DGRC has assembled a large (over 1,000,000 and expanding) inventory of DNA clones through the generosity of individual community members, the BDGP, and the CuraGen Corporation. The collection includes transformation vectors, cDNA clones, and the CuraGen yeast two-hybrid collection. Whenever possible, we will distribute DNAs as spots dried onto FTA filter papers (Whatman). We do not have the resources to distribute ALL collections as complete sets.
Searching for Clones
You can enter the ID for your clone or associated gene in the product search field above. If you are looking for clones in a particular collection, use the Collections menu to the left for any additional tips on finding clones in that collection.
