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. 1999 Nov;67(11):6221-4.
doi: 10.1128/IAI.67.11.6221-6224.1999.

Differential expression of caprine beta-defensins in digestive and respiratory tissues

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Differential expression of caprine beta-defensins in digestive and respiratory tissues

C Zhao et al. Infect Immun. 1999 Nov.

Abstract

We identified two novel beta-defensin precursors, preproGBD-1 and preproGBD-2, in the tissues of a goat. Although the precursors were identical in 96.8% of their bases and 88.2% (60 of 68) of their amino acids, preproGBD-1 was expressed principally in the tongue and respiratory tract, whereas preproGBD-2 expression predominated throughout the intestine. These findings exemplify the phenomenon of tissue-specific expression in a family of host defense peptides that arose before the avian and mammalian lineages diverged.

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Figures

FIG. 1
FIG. 1
cDNA sequences of GBD-1 and GBD-2. The nucleotide and translated sequences of GBD-1 are shown in full. The nucleotide sequence of GBD-2 (311 bp) is shown above that of GBD-1 only where the sequence is different. Amino acids of GBD-2 that differ from those in GBD-1 are shown below the complete GBD-1 sequence. The putative signal sequence is underlined. aa, amino acids.
FIG. 2
FIG. 2
Tissue expression of GBDs. Expression in various tissues was examined by RT-PCR. GBD expression after 25, 30, and 35 PCR cycles is shown, and β-actin expression after 35 cycles is shown. The primers (GBD, P3 and P4; β-actin, P5 and P6) are described in the text. Lanes: M, molecular size markers; 1, bone marrow; 2, mandibular node; 3, mediastinal node; 4, spleen; 5, thymus; 6, pericardium; 7, tongue; 8, mandibular salivary gland; 9, parotid salivary gland; 10, sublingual salivary gland; 11, tonsils; 12, esophagus; 13, reticulum stomach; 14, omasum stomach; 15, abomasum stomach; 16, duodenum; 17, jejunum; 18, ileum; 19, proximal colon; 20, medial colon; 21, distal colon; 22, rectum; 23, anal area; 24, pancreas; 25, liver; 26, gallbladder; 27, renal medulla; 28, renal cortex; 29, bladder-ureter; 30, penis; 31, testis; 32, prostate; 33, epididymis; 34, nasal turbinate; 35, trachea; 36, bronchus; 37, lung; 38, eye; 39, ileocecal node.
FIG. 3
FIG. 3
Northern blot analysis. Total RNA prepared from 39 different goat tissues was probed with the 216-bp PCR product amplified by P3 and P4. The 32P-labeled probe detected both GBD-1 and GBD-2. See the legend to Fig. 2 for identification of lanes.
FIG. 4
FIG. 4
Differential expression of GBD-1 and GBD-2. Differential expression in selected GBD-expressing goat tissues was examined by RT-PCR. The specific primers for GBD-1 were P8 and P9 at an annealing temperature of 58°C. The specific primers for GBD-2 were P10 and P11 at an annealing temperature of 52°C. Additional details are provided in the text. See the legend to Fig. 2 for identification of lanes.
FIG. 5
FIG. 5
Sequences of BDs. References for these peptides are as follows: human BD-1 (2), rhesus monkey BD-1 (14), rat BD-1 (19), mouse BD-1 (11), mouse BD-2 (18), human BD-2 (7), rat BD-2 (13), sheep BD-1 (9, 12), sheep BD-2 (9, 11), cow LAP (22), cow TAP (4), cow EBD (25), cow BNBD4 (28), pig BD-1 (30), chicken (CHIC) GAL-1 (3), chicken GAL-1A (unpublished data); turkey BD-1 (3), turkey BD-2 (3).
FIG. 6
FIG. 6
Dendrogram. The prepropeptide sequences shown in Fig. 5 were analyzed by the Clustal program (IntelliGenetics). CHIC, chicken.

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