A dolphin peripheral blood leukocyte cDNA microarray for studies of immune function and stress reactions.
A microarray focused on stress response and immune function genes of the bottlenosed dolphin has been developed. Random expressed sequence tags (ESTs) were isolated and sequenced from two dolphin peripheral blood leukocyte (PBL) cDNA libraries biased towards T- and B-cell gene expression by stimulation with IL-2 and LPS, respectively. A total of 2784 clones were sequenced and contig analysis yielded 1343 unigenes (archived and annotated at ). In addition, 52 dolphin genes known to be important in innate and adaptive immune function and stress responses of terrestrial mammals were specifically targeted, cloned and added to the unigene collection. The set of dolphin sequences printed on a cDNA microarray comprised the 1343 unigenes, the 52 targeted genes and 2305 randomly selected (but unsequenced) EST clones. This set was printed in duplicate spots, side by side, and in two replicates per slide, such that the total number of features per microarray slide was 19,200, including controls. The dolphin arrays were validated and transcriptomic profiles were generated using PBL from a wild dolphin, a captive dolphin and dolphin skin cells. The results demonstrate that the array is a reproducible and informative tool for assessing differential gene expression in dolphin PBL and in other tissues.
Mancia
Annalaura
A
Marine Biomedicine and Environmental Science Center, Medical University of South Carolina, Hollings Marine Laboratory, 331 Ft. Johnson Road, Charleston, SC 29412, USA. mancia@musc.edu
Lundqvist
Mats L
ML
Romano
Tracy A
TA
Peden-Adams
Margie M
MM
Fair
Patricia A
PA
Kindy
Mark S
MS
Ellis
Blake C
BC
Gattoni-Celli
Sebastiano
S
McKillen
David J
DJ
Trent
Harold F
HF
Chen
Yian Ann
YA
Almeida
Jonas S
JS
Gross
Paul S
PS
Chapman
Robert W
RW
Warr
Gregory W
GW
D000818:Animals; D050681:Bottle-Nosed Dolphin / Q000235:genetics* / Q000276:immunology; D016000:Cluster Analysis; D004847:Epithelial Cells / Q000378:metabolism; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D015723:Gene Library; D007107:Immune System / Q000378:metabolism; D007109:Immunity / Q000235:genetics / Q000502:physiology; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015203:Reproducibility of Results; D013312:Stress, Physiological / Q000503:physiopathology