Individual ESTs may be provided from any of the clone sets, but clones from the Neurospora and Aspergillus libraries are available only by special arrangement.
Other than the Unigene sets, only individual clones will be released.
Strain B3501
Cryptococcus neoformans strain B3501, serotype D was kindly provided by Dr. J. Kwon-Chung
(National Institutes of Health, Bethesda, MD).
C. neoformans yeast cells were cultured in yeast peptone dextrose (YPD) broth at 30°C with shaking for 16 hours.
The cDNA library was constructed by synthesizing cDNA from poly(A)+ RNA using the cDNA Synthesis Kit
(Stratagene, La Jolla, CA) according to the manufacturer's instructions and directionally ligated
into predigested Uni-ZAP XR (Stratagene) and packaged with Gigapack III Gold packaging extracts.
For sequencing, approximately 0.2 ug DNA was used with 20 pM Universal M13 forward primer (5'-TGTAAAACGACGGCCAGT3') or T3 primer (5'-CGAAATTAACCCTCACTAAAG3') The EST database was constructed as follows. The sequences using the Universal forward primer were designated 3' ESTs and given a .f1 suffix while reactions generated using T3 primers were designated 5' ESTs and given a .r1 suffix .
The clones were grown in LB with 50ug/m Ampicillin and 20 ug/ml Kanamycin Host strain is SOLR which is KanR. The vector is pBluescript SK- which is AmpR Cloning was directional into the EcoRI (5'end) and XhoI (3' end) sites
Strain H99
The FGSC received 2 separate EST sets for Neurospora.
The primary EST sets from the UNM and Dartmouth/Oklahoma programs are no longer available. To the extent that clones from the primary sets are in the unigene plates, we will make them available.
Unigene Sets
The Neurospora crassa unigene library was derived from the N. crassa morning and evening libraries (Zhu et al. 2001).
The unigene clone designation refers to the plate (UA-UL) and well.
The contig number refers to the contigs established from the ESTs of the morning and evening libraries.
For more information on the homology (including accession numbers and P/E values) see
http://www.dartmouth.edu/~jdunlap/ESTproject/ESTtable.html
or Nowrousian M, Duffield GE, Loros JJ, Dunlap JC
(2003)Genetics 164: 923-933
More info about the Dartmouth Unigene set (a microsoft Exel File)
The FGSC also holds an internally non-redundant EST set from Drs. M.A. Nelson and D. Natvig
of the University of New Mexico.
This set is comprised of 1784 clones in twenty 96-well plates.
More information is available about the UNM Unigene set (a text file)
Last modified 12/02/04 KMC