Generation and mapping of expressed sequence tags from virus-infected swine macrophages.
In an effort to identify genes that have a major effect on macrophage function during viral infection, we employed differential display reverse transcription (DDRT)-PCR to capture expressed sequence tags (ESTs) of swine alveolar macrophages infected by the porcine reproductive and respiratory syndrome virus (PRRSV). Sequence analyses showed that approximately 60% of these ESTs had significant similarity (> or =93%) to known pig ESTs or genes or matched sequences from other species with homology > or =80%. To determine chromosomal localization, PCR-based mapping was performed across either swine somatic cell hybrid or radiation hybrid panels. A total of 48 porcine viral response ESTs were mapped via the swine somatic cell panel or the INRA-Minnesota porcine Radiation Hybrid (IMpRH) panel (LOD > 6.0). Northern blot analyses confirmed PRRSV-induced altered transcript expression for several ESTs, including a 2'-5' oligoadenylate synthetase and a putative dual-specificity phosphatase. These virus-response ESTs represent good candidate genes for understanding PRRSV pathogenesis and for dissecting host genes which may have major effect on disease resistance.
Wang
C
C
Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St. Paul 55108, USA.
Hawken
R J
RJ
Larson
E
E
Zhang
X
X
Alexander
L
L
Rutherford
M S
MS
D015088:2',5'-Oligoadenylate Synthetase / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D017931:DNA Primers; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016676:Macrophages, Alveolar / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019318:Porcine Reproductive and Respiratory Syndrome / Q000503:physiopathology* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000472:pathogenicity*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013552:Swine
D017931:DNA Primers; D015088:2',5'-Oligoadenylate Synthetase