Dictyostelium discoideum

Sexual cDNA library

"Sexually competent cells"

• The cells are cultured in a liquid medium in the dark, and are competent for fusion with opposite mating type cells.
• They are roughly equivalent to growth phase cells.

Library construction

- done by H. Shimizu using the Super Script Plasmid System (GIBCO/BRL) -
1. mRNA preparation from sexually fusion-competent KAX3 cells
2. oligo-dT primed cDNA synthesis with NotI and SalI linkers on ends
3. Size fractionation to yield two sublibraries, FC and FCL (long)
4. Ligation into pSPORT1 vector

Sublibrary	Size	Mean insert length
FC	> 350,000	1.0 Kb
FCL	>10,000	1.6 Kb

Working team (* indicates previous member)

- Special thanks to Mr. H. Mizuno (Graduate School Institute of Biological Sciences, University of Tsukuba) for his powerful assistance in computer programming.

• Database constsruction and management
  - Hideko Urushihara
  - Yoshihiro Ugawa (AFFRC)
• Nucleotide sequencing
  - Katsuya Suzuki (KS)*
  - Hajime Shimizu (HS)*
  - Asako Kobayashi(AK) *
• Functional analysis
  - Miho Iijima
  - Tomoko Kishimoto*
  - Yukio Aoyama*

Clone and sequence information

We attempt to sequence the whole library but sequences were read by a single pass from 3'and 5'.

• Nucleotide sequences (csm.fasta)
• Clone information
• Search programs (FASTA, BLAST)
• WAIS key word search
  

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For comments or questions, please contact to H. Urushihara