---
id: "edwards-2022-fowlpox"
title: "Genome Sequence of Fowlpox Virus-Integrated Reticuloendotheliosis Virus from a Rio Grande Wild Turkey (Meleagris gallopavo intermedia)"
authors:
  - "Bianca Willis"
  - "Camille Trautman"
  - "Faith Cox"
  - "Tiffany Lujan"
  - "Jason Hardin"
  - "Robert Dittmar"
  - "Camila Romano"
  - "Jeff Brady"
  - "Dustin Edwards"
venue: "Microbiology Resource Announcements"
year: 2022
date: "2022-06-16"
doi: "10.1128/mra.00174-22"
url: "/research/publications/10-1128-mra-00174-22/"
pdf: "/research/publications/10-1128-mra-00174-22/dustin-edwards-10-1128-mra-00174-22.pdf"
pmc: "https://pmc.ncbi.nlm.nih.gov/articles/PMC9202434/"
openAccess: true
license: "cc-by"
accessions:
  - "genbank:OL857287"
  - "sra:SRX11820217"
citedBy: 3
citedBySource: "OpenAlex, read 2026-09-12"
---
# Genome Sequence of Fowlpox Virus-Integrated Reticuloendotheliosis Virus from a Rio Grande Wild Turkey (Meleagris gallopavo intermedia)

A reticuloendotheliosis virus genome found inserted inside a fowlpox virus from a wild turkey in Gillespie County, Texas.

## Abstract

We report the genome sequence of a nearly intact reticuloendotheliosis virus (REV) insertion within a field strain of fowlpox virus from a Rio Grande wild turkey in Gillespie County, TX. The proviral REV genome comprises 7,943 bp and contains partial long terminal repeats.

## Full text

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Genome Sequence of Fowlpox Virus-Integrated
Reticuloendotheliosis Virus from a Rio Grande Wild Turkey
(Meleagris gallopavo intermedia)
Bianca Willis,a,e Camille Trautman,a Faith Cox,a Tiffany Lujan,a,e Jason Hardin,b Robert Dittmar,c† Camila Romano,d Jeff Brady,e
Dustin Edwardsa
aDepartment of Biological Sciences, Tarleton State University, Stephenville, Texas, USA
bTexas Parks and Wildlife Department, Buffalo, Texas, USA
cTexas Parks and Wildlife Department, Kerrville, Texas, USA
dInstituto de Medicina Tropical de São Paulo e Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil
eTexas A&M AgriLife Research, Stephenville, Texas, USA
ABSTRACT We report the genome sequence of a nearly intact reticuloendotheliosis
virus (REV) insertion within a field strain of fowlpox virus from a Rio Grande wild turkey in
Gillespie County, TX. The proviral REV genome comprises 7,943 bp and contains partial
long terminal repeats.
Fowlpox virus (FWPV) is a double-stranded DNA virus within the family Poxviridae that
infects multiple avian species, including domestic poultry and wild birds (1). FWPV has
two presentations: the cutaneous or dry form, in which lesions form on the nonfeathered
areas of the legs, head, and body (2), and the diphtheritic or wet form, in which lesions
form in the mucous membranes of the mouth and respiratory tract (2). Several FWPV vac-
cine and field strains contain integrated partial segments, or near or fully intact genomes,
of the avian gammaretrovirus, reticuloendotheliosis virus (REV) (3, 4). In February 2018, a
landowner in Gillespie County, TX, reported a Rio Grande wild turkey (Meleagris gallopavo
intermedia) with pox-like lesions on the nonfeathered areas of the head and neck. A Texas
Parks and Wildlife Department veterinarian collected lesion, liver, and blood samples.
Histopathology gross findings by the Texas A&M Veterinary Medical Diagnostic Laboratory
(College Station, TX) were compatible with the dry and wet forms of FWPV. DNA was
extracted by excising 5-mm2 sections from a lesion, transferring them into 75
ml HotSHOT
alkaline lysis reagent, and further homogenizing them by pestle. The sample was incubated
at 95°C for 15 min and then cooled, and 75
mL of HotSHOT neutralization solution was
added (5). To determine whether FWPV field isolate TSU-1029 contained an REV proviral ge-
nome insertion, PCR using Hot Start Taq 2 master mix (NEB; M0496L) and the heterol-
ogous primers REV env 7F and FPV 203 4R produced an appropriately sized 740-bp product
(6), which was sequenced using the Sanger method. A BLASTn (https://blast.ncbi.nlm.nih
.gov/) (7) query of the sequence against the nonredundant/nucleotide (nr/nt) database
using MegaBLAST produced an alignment to the integration site of a previously sequenced
REV-integrated fowlpox virus (GenBank accession number AF006064) (8).
A genomic sequencing library was prepared using KAPA HiFi HotStart ReadyMix (Roche;
07958927001) with the primers TR-1 and TR-2 (9), containing Illumina adapters to amplify
the whole REV proviral insertion, flanked by the FWPV open reading frames (ORFs) 201 and
203. The resolved PCR product was excised, tagmented using the Nextera XT DNA library
prep kit (Illumina; FC-131-1096), pooled, and sequenced using the MiSeq reagent kit v3 (600
cycles) (Illumina; MS-102-3003). Sequencing produced 4,238 total paired-end reads with a
read length of 300 bp. The primer sequences and low-quality and short (,30 bp) reads
were trimmed. A consensus sequence was assembled from the reference sequence found
Editor Jelle Matthijnssens, KU Leuven
Copyright © 2022 Willis et al. This is an open-
access article distributed under the terms of
the Creative Commons Attribution 4.0
International license.
Address correspondence to Dustin Edwards,
dcedwards@tarleton.edu.
The authors declare no conflict of interest.
†Deceased.
Received 21 February 2022
Accepted 29 April 2022
Published 16 May 2022
June 2022 Volume 11 Issue 6 10.1128/mra.00174-22 1
GENOME SEQUENCES
Downloaded from https://journals.asm.org/journal/mra on 26 July 2026 by 156.146.253.207.

under GenBank accession number AF246698.2 (10) and the sequence reads using the CLC
Genomics Workbench v6 assemble to reference tool (mismatch cost = 1; indels cost = 2;
minimum fraction of similarity between reads and reference = 90%). A total of 92.4% of
trimmed reads were mapped (average coverage, 50). Genome annotation was performed
using SeqBuilder Pro v17.2.1 to assign open reading frames and determine the long terminal
repeats (LTRs). For analysis of the REV sequence, the annotated reference and consensus ge-
nome sequences were aligned using ClustalW in MEGA v11.011 (11), and the FWPV ORF 201
and 203 sequences were manually identified and trimmed. All tools were run with default
parameters unless indicated. The proviral REV genome comprises 7,943 bp and has a 52.4%
G1C content. BLASTn (nr/nt) searches displayed 7,942/7,943 and 7,937/7,943 nucleotide
identities to nearly intact REV proviral genomes within the genomes of FWPV-SD15-670.2
(MH734528) and FWPV-MN00.2 (MH709124) from Merriam’s wild turkeys (Meleagris gallo-
pavo merriami) (3). Compared to the full-length REV strain 104865 (KJ756349), this genome
shared 7,940/7,943 nucleotide identities with single nucleotide polymorphisms at positions
3 and 7646 and an insertion at position 7. In addition, a 23-bp repeat was deleted at the
59 end, and the 39 long terminal repeat was truncated to 222 bp, producing partial LTRs.
Data availability. The sequence has been deposited at GenBank under accession
number OL857287 and the raw reads under SRA accession number SRX11820217.
ACKNOWLEDGMENTS
We thank Texas Parks and Wildlife Department Upland Game Bird biologists for the
wild turkey sample. Support for this research was provided by Tarleton State University and
the College of Science and Technology for Undergraduate Research Assistantships and
Student Research Grants.
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Announcement Microbiology Resource Announcements
June 2022 Volume 11 Issue 6 10.1128/mra.00174-22 2
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