A strain of highly pathogenic porcine reproductive and respiratory syndrome virus: genomic characterization, pathogenicity, and construction of an infectious full-length cDNA clone

2021 ◽  
Author(s):  
Shuangyun Wang ◽  
Yanling Liu ◽  
Linyang Yu ◽  
Tairun Liang ◽  
Pengfei Zhang ◽  
...  
Keyword(s):  
Cdna Clone ◽  
Full Length ◽  
Respiratory Syndrome Virus ◽  
Highly Pathogenic ◽  
Full Length Cdna ◽  
Genomic Characterization

scholarly journals Infectious Transcripts from Cloned Genome-Length cDNA of Porcine Reproductive and Respiratory Syndrome Virus

1998 ◽  
Vol 72 (1) ◽  
pp. 380-387 ◽  
Author(s):  
J. J. M. Meulenberg ◽  
J. N. A. Bos-de Ruijter ◽  
R. van de Graaf ◽  
G. Wensvoort ◽  
R. J. M. Moormann
Keyword(s):  
Cdna Clone ◽  
De Novo ◽  
Infectious Clone ◽  
Full Length ◽  
Respiratory Syndrome Virus ◽  
Cdna Clones ◽  
Genome Length ◽  
Full Length Cdna ◽  
Lung Macrophages ◽  
Growth Properties

ABSTRACT The 5′-terminal end of the genomic RNA of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was determined. To construct full-length cDNA clones, the 5′-terminal sequence was ligated to cDNA clones covering the complete genome of LV. When RNA that was transcribed in vitro from these full-length cDNA clones was transfected into BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages or CL2621 cells, no infectious virus was produced due to the poor transfection efficiency of these cells. The growth properties of the viruses produced by BHK-21 cells transfected with infectious transcripts of LV cDNA resembled the growth properties of the parental virus from which the cDNA was derived. Two nucleotide changes leading to a unique PacI restriction site directly downstream of the ORF7 gene were introduced in the genome-length cDNA clone. The virus recovered from this mutated cDNA clone retained the PacI site, which confirmed the de novo generation of infectious LV from cloned cDNA. These results indicate that the infectious clone of LV enables us to mutagenize the viral genome at specific sites and that it will therefore be useful for detailed molecular characterization of the virus, as well as for the development of a safe and effective live vaccine for use in pigs.

scholarly journals Generation of an Infectious Clone of VR-2332, a Highly Virulent North American-Type Isolate of Porcine Reproductive and Respiratory Syndrome Virus

2003 ◽  
Vol 77 (6) ◽  
pp. 3702-3711 ◽  
Author(s):  
H. S. Nielsen ◽  
G. Liu ◽  
J. Nielsen ◽  
M. B. Oleksiewicz ◽  
A. Bøtner ◽  
...  
Keyword(s):  
Growth Kinetics ◽  
Cdna Clone ◽  
North American ◽  
Molecular Mechanisms ◽  
Full Length ◽  
Respiratory Syndrome Virus ◽  
Full Length Cdna ◽  
Kinetics Of ◽  
Highly Virulent

ABSTRACT A full-length cDNA clone of the prototypical North American porcine reproductive and respiratory syndrome virus (PRRSV) isolate VR-2332 was assembled in the plasmid vector pOK12. To rescue infectious virus, capped RNA was transcribed in vitro from the pOK12 clone and transfected into BHK-21C cells. The supernatant from transfected monolayers were serially passaged on Marc-145 cells and porcine pulmonary alveolar macrophages. Infectious PRRSV was recovered on Marc-145 cells as well as porcine pulmonary macrophages; thus, the cloned virus exhibited the same cell tropism as the parental VR-2332 strain. However, the cloned virus was clearly distinguishable from the parental VR-2332 strain by an engineered marker, a BstZ17I restriction site. The full-length cDNA clone had 11 nucleotide changes, 2 of which affected coding, compared to the parental VR-2332 strain. Additionally, the transcribed RNA had an extra G at the 5′ end. To examine whether these changes influenced viral replication, we examined the growth kinetics of the cloned virus in vitro. In Marc-145 cells, the growth kinetics of the cloned virus reflected those of the parental isolate, even though the titers of the cloned virus were consistently slightly lower. In experimentally infected 5.5-week-old pigs, the cloned virus produced blue discoloration of the ears, a classical clinical symptom of PRRSV. Also, the seroconversion kinetics of pigs infected with the cloned virus and VR-2332 were very similar. Hence, virus derived from the full-length cDNA clone appeared to recapitulate the biological properties of the highly virulent parental VR-2332 strain. This is the first report of an infectious cDNA clone based on American-type PRRSV. The availability of this cDNA clone will allow examination of the molecular mechanisms behind PRRSV virulence and attenuation, which might in turn allow the production of second-generation, genetically engineered PRRSV vaccines.

Construction of an infectious full-length cDNA clone of potato aucuba mosaic virus

2021 ◽  
Vol 166 (5) ◽  
pp. 1427-1431
Author(s):  
Buyang Chen ◽  
Qi Lin ◽  
Yueyan Yin ◽  
Liangliang Jiang ◽  
Fang Wang ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Cdna Clone ◽  
Full Length ◽  
Full Length Cdna ◽  
Aucuba Mosaic

Construction of an infectious full-length cDNA clone of Chrysanthemum virus B

2008 ◽  
Vol 74 (6) ◽  
pp. 434-437 ◽  
Author(s):  
Atsushi Ohkawa ◽  
Noriko Ishikawa-Suehiro ◽  
Seiichi Okuda ◽  
Tomohide Natsuaki
Keyword(s):  
Cdna Clone ◽  
Full Length ◽  
Full Length Cdna

scholarly journals Nucleotide sequence of a full-length cDNA clone encoding preproparathyroid hormone from pig and rat

1987 ◽  
Vol 15 (16) ◽  
pp. 6740-6740 ◽  
Author(s):  
Hans-Jürgen Schmelzer ◽  
Gerhard Gross ◽  
Georg Widera ◽  
Hubert Mayer
Keyword(s):  
Nucleotide Sequence ◽  
Cdna Clone ◽  
Full Length ◽  
Full Length Cdna

scholarly journals Beta-centractin: characterization and distribution of a new member of the centractin family of actin-related proteins.

1994 ◽  
Vol 5 (12) ◽  
pp. 1301-1310 ◽  
Author(s):  
S W Clark ◽  
O Staub ◽  
I B Clark ◽  
E L Holzbaur ◽  
B M Paschal ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cdna Clone ◽  
Expressed Sequence Tag ◽  
Full Length ◽  
Northern Analysis ◽  
Related Protein ◽  
Two Dimensional Gel Electrophoresis ◽  
Full Length Cdna ◽  
Dynactin Complex ◽  
Expressed Sequence

An examination of human-expressed sequence tags indicated the existence of an isoform of centractin, an actin-related protein localized to microtubule-associated structures. Using one of these tags, we isolated and determined the nucleotide sequence of a full-length cDNA clone. The protein encoded represents the first example of multiple isoforms of an actin-related protein in a single organism. Northern analysis using centractin-specific probes revealed three species of mRNA in HeLa cells that could encode centractin isoforms. One mRNA encodes the previously-identified centractin (now referred to as alpha-centractin). The full-length cDNA clone isolated using the expressed sequence tag encodes a new member of the centractin family, beta-centractin. A probe specific for alpha-centractin hybridized to the third species of mRNA observed (referred to as gamma-centractin). Comparisons of Northern blots of human tissues indicated that alpha-centractin and beta-centractin mRNAs are equally distributed in all populations of mRNA examined, whereas the expression of gamma-centractin appears to be tissue specific. The amino acid sequence of beta-centractin, deduced from the cDNA, indicates a 91% identity with alpha-centractin, increasing to 96% similarity when conservative amino acid changes are taken into account. As antibodies previously raised against alpha-centractin reacted only poorly with beta-centractin, new antibodies were produced and combined with two-dimensional gel electrophoresis to discriminate the two isoforms. Using this system, the subcellular distribution of the alpha- and beta-isoforms were determined. Both isoforms were found predominantly in the cytosolic fraction as a part of a previously identified 20S complex (referred to as the dynactin complex) with no evidence for a free pool of either isoform. The isoforms were found in a constant ratio of approximately 15:1 (alpha:beta) in the dynactin complex.

scholarly journals Synthesis of an infectious full-length cDNA clone of rice yellow mottle virus andmutagenesis of the coat protein

Virology ◽  
1995 ◽  
Vol 206 (1) ◽  
pp. 108-115 ◽  
Author(s):  
C. Brugidou ◽  
C. Holt ◽  
M. Ngon A Yassi ◽  
S. Zhang ◽  
R. Beachy ◽  
...  
Keyword(s):  
Coat Protein ◽  
Cdna Clone ◽  
Full Length ◽  
Mottle Virus ◽  
Rice Yellow Mottle Virus ◽  
Full Length Cdna ◽  
Yellow Mottle

Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants

2010 ◽  
Vol 142 (1-2) ◽  
pp. 3-12 ◽  
Author(s):  
Katrin Mischkale ◽  
Ilona Reimann ◽  
J. Zemke ◽  
P. König ◽  
Martin Beer
Keyword(s):  
Cdna Clone ◽  
Full Length ◽  
Full Length Cdna ◽  
Genotype 2
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