Aspergillus gene libraries

24 h developmental cDNA library from Aspergillus nidulans constructed by R.Aramayo

It was deposited to the FGSC in 1995 and maintained as phage lysate in SM at -80 with one freeze-thaw cycle to aliquot samples.

The developmental poly(A)+ mRNA used to construct this library was prepared as follows: conidia from the strain FGSC A26 (biA1) were germinated in a liquid medium and allowed to grow 18 h. Mycelia was harvested on top of filter paper and placed onto appropriately supplemented minimal medium. Induced mycelia were then allowed to develop for 24 h prior to harvest and mRNA extraction. Poly(A)+ mRNA was prepared using standard procedures. The cDNA library was constructed using the lambda ZAPcDNA TM+ kit lot # UC105. The primary library contained approximately 100,000 clones. After plating this primary library, the phage were harvested and stored at -70øC as suggested by Maniatis. This library has not been amplified. Genes expressed as early as 10 h have been cloned from this library.

The Aspergillus gene libraries below should be considered ARCHIVAL.

All of them were deposited at the FGSC more than ten years ago.

Ordered A. nidulans  Genomic DNA libraries

pWE15 and pLORIST2 genomic cosmid libraries

The FGSC distributed these cosmid libraries (30 microtiter plates each) either separately, or more typically, together. Together, the two libraries give good coverage of the A. nidulans genome and form the basis of the chromosome specific and minimal libraries.

Vector Maps:
 pWE15
 pLORIST2
a list of identified clones in the pWE15 and pLORIST2 libraries

Chromosome specific libraries
All clones from the pWE15 and pLORIST2 libraries showing hybridization to chromosome-specific probes were reisolated to microtiter plates according their chromosome linkage (Brody et al. 1991. Nucleic Acids Res.19:3105-3109). A total of 38 chromosome specific plates resulted. These were sent as a complete set or as individual chromosome subsets.

Minimal Compressed library from A. nidulans.
The minimal, ordered, compressed libraries of A. nidulans are described in Prade et al. PNAS 94:14564-14569 (1997). These libraries were picked from the pWE15 and pLORIST2 sets at the FGSC and were distributed in 16 96 well microtitre plates.

Broad (WICGR) Institute fosmid library

Individual clones from the A. nidulans sequencing program at the Broad Institute are available.

This spreadsheet lists the well identities.

Unordered Genomic DNA libraries

A. fumigatus libraries from G. May/ J. Yu.
We have received amplified aliquots of two genomic libraries for Aspergillus fumigatus. The libraries were produced in Greg May's lab at M.D. Anderson Cancer Center and are described in Arch Microbiol (2004) 182: 346–353. They were amplified by Jaehyuk Yu at the University of Wisconsin. The two libraries are in the vector pRG3-AMA-NotI. We titred them at the FGSC and found that the 4-9Kb library contained 7.5 X 109 cfu/ ml and the 9 Kb+ library contained 3.6 X 109 cfu/ml. We can distribute them as 250 ul aliquots of E. coli cells frozen in glycerol.

A. nidulans libraries

These were sent as a small aliquot of plasmid DNA suitable for transforming E. coli for amplification. These libraries have been amplified at the FGSC.
See: Polarity-Defective Mutants of Aspergillus nidulans, N.Osherov, J.Mathew, and G.S. May Fungal Genet Biol, 31: 181-188

Primers for sequencing inserts in these AMA vectors directly

cDNA libraries

Lambda GT10 cDNA library
Prepared from hyphae of an A. nidulans strain called R153 (wA3 pyroA4) grown on yeast extract glucose medium.
See: Cell. 1988 Apr 22;53(2):237-44. This was sent as a sample of packaged phage.

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