An annotated cDNA library of juvenile Euprymna scolopes with and without colonization by the symbiont Vibrio fischeri.
BACKGROUND
Biologists are becoming increasingly aware that the interaction of animals, including humans, with their coevolved bacterial partners is essential for health. This growing awareness has been a driving force for the development of models for the study of beneficial animal-bacterial interactions. In the squid-vibrio model, symbiotic Vibrio fischeri induce dramatic developmental changes in the light organ of host Euprymna scolopes over the first hours to days of their partnership. We report here the creation of a juvenile light-organ specific EST database.


RESULTS
We generated eleven cDNA libraries from the light organ of E. scolopes at developmentally significant time points with and without colonization by V. fischeri. Single pass 3' sequencing efforts generated 42,564 expressed sequence tags (ESTs) of which 35,421 passed our quality criteria and were then clustered via the UIcluster program into 13,962 nonredundant sequences. The cDNA clones representing these nonredundant sequences were sequenced from the 5' end of the vector and 58% of these resulting sequences overlapped significantly with the associated 3' sequence to generate 8,067 contigs with an average sequence length of 1,065 bp. All sequences were annotated with BLASTX (E-value < -03) and Gene Ontology (GO).


CONCLUSIONS
Both the number of ESTs generated from each library and GO categorizations are reflective of the activity state of the light organ during these early stages of symbiosis. Future analyses of the sequences identified in these libraries promise to provide valuable information not only about pathways involved in colonization and early development of the squid light organ, but also about pathways conserved in response to bacterial colonization across the animal kingdom.
Chun
Carlene K
CK
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA. cchun@wisc.edu
Scheetz
Todd E
TE
Bonaldo
Maria de Fatima
Mde F
Brown
Bartley
B
Clemens
Anik
A
Crookes-Goodson
Wendy J
WJ
Crouch
Keith
K
DeMartini
Tad
T
Eyestone
Mari
M
Goodson
Michael S
MS
Janssens
Bernadette
B
Kimbell
Jennifer L
JL
Koropatnick
Tanya A
TA
Kucaba
Tamara
T
Smith
Christina
C
Stewart
Jennifer J
JJ
Tong
Deyan
D
Troll
Joshua V
JV
Webster
Sarahrose
S
Winhall-Rice
Jane
J
Yap
Cory
C
Casavant
Thomas L
TL
McFall-Ngai
Margaret J
MJ
Soares
M Bento
MB
D048908:Aliivibrio fischeri / Q000254:growth & development*; D000818:Animals; D001483:Base Sequence; D049832:Decapodiformes / Q000235:genetics* / Q000254:growth & development / Q000382:microbiology; D020224:Expressed Sequence Tags; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013559:Symbiosis / Q000502:physiology*