scholarly journals Apple Russet Ring and Apple Green Crinkle Diseases: Fulfillment of Koch’s Postulates by Virome Analysis, Amplification of Full-Length cDNA of Viral Genomes, in vitro Transcription of Infectious Viral RNAs, and Reproduction of Symptoms on Fruits of Apple Trees Inoculated With Viral RNAs

2020 ◽  
Vol 11 ◽  
Author(s):  
Chunjiang Li ◽  
Hajime Yaegashi ◽  
Ryusuke Kishigami ◽  
Ayaka Kawakubo ◽  
Noriko Yamagishi ◽  
...  
Keyword(s):  
In Vitro Transcription ◽  
Full Length ◽  
Apple Trees ◽  
Koch’S Postulates ◽  
Viral Rnas ◽  
Viral Genomes ◽  
Full Length Cdna ◽  
Koch's Postulates

In vitro transcription and translation of a full-length cDNA coding for the human ribosomal protein Lia

1993 ◽  
Vol 4 (1) ◽  
pp. 83-88
Author(s):  
Giulia Russo ◽  
Sandro De Falco ◽  
Antonietta Angiolillo ◽  
Concetta Pietropaolo ◽  
Socio A. Ruffo
Keyword(s):  
Ribosomal Protein ◽  
In Vitro Transcription ◽  
Full Length ◽  
Full Length Cdna ◽  
Human Ribosomal Protein

Establishment of Full-length cDNA Clones and an Efficient Oral Infection Model for Feline Coronavirus in Cats

2021 ◽  
Author(s):  
Gang Wang ◽  
Guangli Hu ◽  
Rui Liang ◽  
Jiale Shi ◽  
Xiuxiu Qiu ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Full Length ◽  
Infection Model ◽  
Type I ◽  
Feline Infectious Peritonitis ◽  
Spike Gene ◽  
Full Length Cdna ◽  
Adult Cats

Feline infectious peritonitis virus (FIPV) is the etiologic agent of feline infectious peritonitis (FIP) and causes fatal disease in cats of almost all ages. Currently, there are no clinically approved drugs or effective vaccines for FIP. Furthermore, the pathogenesis of FIP is still not fully understood. There is an urgent need for an effective infection model of feline infectious peritonitis induced by FIPV. Here, we constructed a field type I FIPV full-length cDNA clone, pBAC-QS, corresponding to the isolated FIPV QS. By replacing the FIPV QS spike gene with the commercially available type II FIPV 79-1146 (79-1146_CA) spike gene, we established and rescued a recombinant virus, designated rQS-79. Moreover, we constructed 79-1146_CA infectious full-length cDNA pBAC-79-1146_CA, corresponding to recombinant FCoV 79-1146_CA (r79-1146_CA). In animal experiments with one- to two-year-old adult cats orally infected with the recombinant virus, rQS-79 induced typical FIP signs and 100% mortality. In contrast to cats infected with rQS-79, cats infected with 79-1146_CA did not show obvious signs. Furthermore, by rechallenging rQS-79 in surviving cats previously infected with 79-1146_CA, we found that there was no protection against rQS-79 with different titers of neutralizing antibodies. However, high titers of neutralizing antibodies may help prolong the cat survival time. Overall, we report the first reverse genetics of virulent recombinant FCoV (causing 100% mortality in adult cats) and attenuated FCoV (causing no mortality in adult cats), which will be powerful tools to study the pathogenesis, antiviral drugs and vaccines for FCoV. Importance Tissue- or cell culture-adapted feline infectious peritonitis virus (FIPV) usually loses pathogenicity. To develop a highly virulent FIPV, we constructed a field isolate type I FIPV full-length clone with the spike gene replaced by the 79-1146 spike gene, corresponding to a virus named rQS-79, which induces high mortality in adult cats. rQS-79 represents the first described reverse genetics system for highly pathogenic FCoV. By further constructing the cell culture-adapted FCoV 79-1146_CA, we obtained infectious clones of virulent and attenuated FCoV. By in vitro and in vivo experiments, we established a model that can serve to study the pathogenic mechanisms of FIPV. Importantly, the wild-type FIPV replicase skeleton of serotype I will greatly facilitate the screening of antiviral drugs, both in vivo and in vitro.

Complete sequence and in vitro expression of a tissue-specific phosphatidylinositol-linked N-CAM isoform from skeletal muscle

Development ◽  
1988 ◽  
Vol 104 (1) ◽  
pp. 165-173 ◽  
Author(s):  
C.H. Barton ◽  
G. Dickson ◽  
H.J. Gower ◽  
L.H. Rowett ◽  
W. Putt ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Amino Acids ◽  
Cell Surface ◽  
Protein Sequence ◽  
Single Copy ◽  
In Vitro Transcription ◽  
Full Length ◽  
Covalent Attachment ◽  
Size Difference

Neural cell adhesion molecules (N-CAMs) are a family of cell surface sialoglycoproteins encoded by a single copy gene. A full-length cDNA clone that encodes a nontransmembrane phosphatidylinositol (PI) linked N-CAM of Mr 125 × 10(3) has been isolated from a human skeletal muscle cDNA library. The deduced protein sequence encodes a polypeptide of 761 amino acids and is highly homologous to the N-CAM isoform in brain of Mr 120 × 10(3). The size difference between the 125 × 10(3). The size difference between the 125 × 10(3) Mr skeletal muscle form and the 120 × 10(3) Mr N-CAM form from brain is accounted for by the insertion of a block of 37 amino acids called MSD1, in the extracellular domain of the muscle form. Transient expression of the human cDNA in COS cells results in cell surface N-CAM expression via a putative covalent attachment to PI-containing phospholipid. Linked in vitro transcription and translation experiments followed by immunoprecipitation with anti-N-CAM antibodies demonstrate that the full-length clone of 761 amino acid coding potential produces a core polypeptide of Mr 110 × 10(3) which is processed by microsomal membranes to yield a 122 × 10(3) Mr species. Taken together, these results demonstrate that the cloned cDNA sequence encodes a lipid-linked, PI-specific phospholipase C releasable surface isoform of N-CAM with core glycopeptide molecular weight corresponding to the authentic muscle 125 × 10(3) Mr N-CAM isoform. This is the first direct correlation of cDNA and deduced protein sequence with a known PI-linked N-CAM isoform from skeletal muscle.

scholarly journals In vitro transcription of infectious RNAs from full-length cDNAs of tobacco mosaic virus

1986 ◽  
Vol 83 (14) ◽  
pp. 5043-5047 ◽  
Author(s):  
T. Meshi ◽  
M. Ishikawa ◽  
F. Motoyoshi ◽  
K. Semba ◽  
Y. Okada
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
In Vitro Transcription ◽  
Full Length

Isolation of a full-length cDNA encoding calreticulin from a PCR library of in vitro zygotes of maize

1996 ◽  
Vol 31 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Thomas Dresselhaus ◽  
Christine Hagel ◽  
Horst L�rz ◽  
Erhard Kranz
Keyword(s):  
Full Length ◽  
Full Length Cdna

scholarly journals Infectious genomic RNA of Rhopalosiphum padi virus transcribed in vitro from a full-length cDNA clone

Virology ◽  
2008 ◽  
Vol 375 (2) ◽  
pp. 401-411 ◽  
Author(s):  
Sandhya Boyapalle ◽  
Randy J. Beckett ◽  
Narinder Pal ◽  
W. Allen Miller ◽  
Bryony C. Bonning
Keyword(s):  
Cdna Clone ◽  
Rhopalosiphum Padi ◽  
Full Length ◽  
Genomic Rna ◽  
Full Length Cdna
Sign in / Sign up

Export Citation Format

Share Document