Construction and characterization of an expressed sequenced tag library for the mosquito vector Armigeres subalbatus

Table 4 Searching Ar. subalbatus EST clusters for immunity-related sequences based on homology to other flies.

Family	Subfamily	asuhem	diroinf	imacbac	brumal	naïve7	naïve14	Total Reads	R Value
AMP	Cecropin	98	65	70	56	41	47	377	0.070988
AMP	*Defensin*	*238*	41	78	16	15	28	*416*	0
AMP	Diptericin	0	2	0	0	0	1	3	0.160764
CASP	CASP	1	1	0	1	1	0	4	0.673041
CAT	CAT	0	1	0	0	0	0	1	0.561495
CLIP	CLIP-CLIP-CLIP-SPH	1	1	1	0	0	1	4	0.670142
CLIP	CLIP-SP	136	33	37	24	22	27	279	0
CLIP	CLIP-SP-CLIP-SP	1	0	0	1	0	0	2	0.668311
CLIP	CLIP-SPH	153	33	55	58	14	47	360	0
CLIP	SP-CLIP	4	0	0	0	0	0	4	0.060745
CLIP	SP-CLIP-SPH	2	0	0	1	0	0	3	0.440816
CLIP	SPH-CLIP	1	0	0	0	0	0	1	0.754901
CLIP	SPH-CLIP-SPH	1	1	0	2	0	1	5	0.576541
CLIP	SP-SPH-CLIP	0	1	0	0	0	0	1	0.561495
CTL	CTL	80	19	27	17	25	17	185	0.000002
FREP	FREP	17	9	22	4	4	8	64	0.000764
GALE	GALE	0	0	2	1	1	2	6	0.206431
GNBP	GNBP	0	1	1	0	0	0	2	0.420516
IAP	IAP	1	1	0	3	0	2	7	0.260929
IMDPATH	IKKg	1	0	0	1	0	0	2	0.668311
LYS	LYSC	122	21	37	19	15	42	256	0
ML	ML	14	10	5	8	9	3	49	0.223609
PGRP	PGRP	9	1	4	2	2	5	23	0.312906
PPO	PPO	1	1	0	1	0	0	3	0.653363
PRDX	GPX	8	2	1	1	0	3	15	0.100553
PRDX	HPX	1	2	1	3	1	0	8	0.446023
PRDX	TPX	7	3	5	14	4	8	41	0.136651
SCR	SCRA	0	7	3	1	2	2	15	0.004735
SCR	SCRB	1	0	0	1	2	0	4	0.26927
SCR	SCRC	1	0	0	0	0	0	1	0.754901
SOD	SOD-Cu-Zn	5	3	3	7	2	2	22	0.798952
SOD	SOD-Mn-Fe	30	5	4	2	4	6	51	0.000052
SPZ	SPZ	0	0	1	0	0	0	1	0.575875
SRPN	SRPN-INHIB	26	8	8	2	8	4	56	0.001248
SRPN	SRPN-nonINHIB	4	2	2	0	3	2	13	0.281764
TEP	TEP	1	1	2	1	1	4	10	0.376665
TOLLPATH	TUBE	0	0	0	0	0	1	1	0.562898
	Totals	965	275	369	247	176	263	2295

Peptide sequences were harvested from ImmunoDB [24] for An. gambiae, Ae. aegypti, and D. melanogaster for each of the following gene families: All 8,020 EST clusters were blasted against a database of these peptides using blastx (e-value cutoff of 1e^-5). BLAST hits were parsed from output files using tcl_blast_parser_123_V017.tcl [55] with a cut-off of 40% match, 1e^-20 normalized e-value, and a minimum match length of 30 residues. Top hits were taken for each significant match and verified manually. Matches then were categorized by family and subfamily according to Waterhouse et al. (2007) (Left column) and the composition (# of compiled sequences) of EST clusters were collated from the assembly according to the library(s) from which ESTs came: asuhem (bacteria-inoculated, hemocyte), diroinf (whole body, Dirofilaria immitis injected), imacbac (bacteria-inoculated whole body), brumal (blood-fed Brugia malayi), n7 and n14 (naïve, 5–7 and 12–14 days of age). Stekel R values [23] were determined for the groupings using IDEG6 [20] and those having a score of 0.001 or better were considered significantly differential, and are flagged with bolded and underlined text for the row.

ISSN: 1471-2164