Infectious in Vitro Transcripts From cDNA Clone of a Japanese Gentian Isolate of Sikte Waterborne Virus, Which Shows Host-specific Low-temperature-preferred Multiplication

2021 ◽  
Author(s):  
Koki Fujisaki ◽  
Chika Tateda ◽  
Yoshiko Abe ◽  
John Jewish A. Dominguez ◽  
Mari Iwai ◽  
...  
Keyword(s):  
Low Temperature ◽  
Virus Strain ◽  
Cdna Clone ◽  
Cdna Clones ◽  
Genomic Rna ◽  
Rna Sequences ◽  
Japanese Gentian ◽  
In Vitro Transcripts ◽  
Waterborne Virus

Abstract Tombusviruses have been identified in several crops, which include gentian virus A (GeVA), in Japanese gentians. In this study, we isolated another tombusvirus, Sikte waterborne virus strain C1 (SWBV-C1) from Japanese gentian. Although SWBV-C1 and GeVA are not closely related among tombusviruses, SWBV-C1, like GeVA, showed host-specific low-temperature-preferred multiplication in gentians and Arabidopsis. The use of in vitro transcripts from full-length cDNA clones of SWBV-C1 genomic RNA as inocula confirmed these properties, which indicates that the identified genomic RNA sequences encode viral factors underlying characteristic SWBV-C1 features.

scholarly journals In vitro expression of human p53 cDNA clones and characterization of the cloned human p53 gene.

1985 ◽  
Vol 5 (8) ◽  
pp. 1887-1893 ◽  
Author(s):  
D Wolf ◽  
Z Laver-Rudich ◽  
V Rotter
Keyword(s):  
Cdna Clone ◽  
P53 Protein ◽  
Cdna Probe ◽  
P53 Gene ◽  
Heteroduplex Analysis ◽  
Cdna Clones ◽  
Free System ◽  
Human Cdna ◽  
Mrna Molecule

The human p53 gene was cloned and characterized by using a battery of p53 DNA clones. A series of human cDNA clones of various sizes and relative localizations to the mRNA molecule were isolated by using the human p53-H14 (2.35-kilobase) cDNA probe which we previously cloned. One such isolate, clone p53-H7 (2.65 kilobases), spans the entire human mature p53 mRNA molecule. Construction of the human cDNA clones in the pSP65 RNA transcription vector facilitated the generation of p53 transcripts by the SP6 bacteriophage RNA polymerase. The p53-specific RNA transcripts obtained without further processing were translated into p53 proteins in a cell-free system. By using this rapid in vitro transcription-translation assay, we found that whereas clone p53-H7 (2.65 kilobases) coded for a mature-sized p53 protein, a shorter cDNA clone, p53-H13 (1.8 kilobases), dictated the synthesis of a smaller-sized p53 protein (45 kilodaltons). The p53 proteins synthesized in vitro immunoprecipitated efficiently with human-specific anti-p53 antibodies. Genomic analysis of human DNA revealed the presence of a single p53 gene residing within two EcoRI fragments. Heteroduplex analysis between the full-length cDNA clone p53-H7 and the cloned p53 gene indicated the presence of seven major exons.

Infectious in vitro transcripts from a cDNA clone of Tobacco mild green mosaic tobamovirus and its biological activity in host and nonhost plants and in their protoplasts

2003 ◽  
Vol 69 (5) ◽  
pp. 335-338 ◽  
Author(s):  
Naoto Morishima ◽  
Takao Ido ◽  
Hiroyuki Hamada ◽  
Eri Yoshimoto ◽  
Hiroyuki Mizumoto ◽  
...  
Keyword(s):  
Biological Activity ◽  
Cdna Clone ◽  
In Vitro Transcripts

Complete genomic RNA sequences of cucumber mosaic virus strain NT9 from Taiwan

1995 ◽  
Vol 140 (10) ◽  
pp. 1841-1847 ◽  
Author(s):  
Y. -H. Hsu ◽  
C. -W. Wu ◽  
B. -Y. Lin ◽  
H. -Y. Chen ◽  
M. -F. Lee ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Strain ◽  
Genomic Rna ◽  
Rna Sequences ◽  
Cucumber Mosaic Virus Strain ◽  
Complete Genomic

Assembly of the Alu domain of the signal recognition particle (SRP): dimerization of the two protein components is required for efficient binding to SRP RNA

1990 ◽  
Vol 10 (2) ◽  
pp. 777-784
Author(s):  
K Strub ◽  
P Walter
Keyword(s):  
Cdna Clone ◽  
Repetitive Sequences ◽  
Signal Recognition Particle ◽  
Secretory Proteins ◽  
Endoplasmic Reticulum Membrane ◽  
Cdna Clones ◽  
Functional Domain ◽  
Signal Recognition ◽  
Srp Rna

The signal recognition particle (SRP), a cytoplasmic ribonucleoprotein, plays an essential role in targeting secretory proteins to the rough endoplasmic reticulum membrane. In addition to the targeting function, SRP contains an elongation arrest or pausing function. This function is carried out by the Alu domain, which consists of two proteins, SRP9 and SRP14, and the portion of SRP (7SL) RNA which is homologous to the Alu family of repetitive sequences. To study the assembly pathway of the components in the Alu domain, we have isolated a cDNA clone of SRP9, in addition to a previously obtained cDNA clone of SRP14. We show that neither SRP9 nor SRP14 alone interacts specifically with SRP RNA. Rather, the presence of both proteins is required for the formation of a stable RNA-protein complex. Furthermore, heterodimerization of SRP9 and SRP14 occurs in the absence of SRP RNA. Since a partially reconstituted SRP lacking SRP9 and SRP14 [SRP(-9/14)] is deficient in the elongation arrest function, it follows from our results that both proteins are required to assemble a functional domain. In addition, SRP9 and SRP14 synthesized in vitro from synthetic mRNAs derived from their cDNA clones restore elongation arrest activity to SRP(-9/14).

In vitro expression of human p53 cDNA clones and characterization of the cloned human p53 gene

1985 ◽  
Vol 5 (8) ◽  
pp. 1887-1893
Author(s):  
D Wolf ◽  
Z Laver-Rudich ◽  
V Rotter
Keyword(s):  
Cdna Clone ◽  
P53 Protein ◽  
Cdna Probe ◽  
P53 Gene ◽  
Heteroduplex Analysis ◽  
Cdna Clones ◽  
Free System ◽  
Human Cdna ◽  
Mrna Molecule

The human p53 gene was cloned and characterized by using a battery of p53 DNA clones. A series of human cDNA clones of various sizes and relative localizations to the mRNA molecule were isolated by using the human p53-H14 (2.35-kilobase) cDNA probe which we previously cloned. One such isolate, clone p53-H7 (2.65 kilobases), spans the entire human mature p53 mRNA molecule. Construction of the human cDNA clones in the pSP65 RNA transcription vector facilitated the generation of p53 transcripts by the SP6 bacteriophage RNA polymerase. The p53-specific RNA transcripts obtained without further processing were translated into p53 proteins in a cell-free system. By using this rapid in vitro transcription-translation assay, we found that whereas clone p53-H7 (2.65 kilobases) coded for a mature-sized p53 protein, a shorter cDNA clone, p53-H13 (1.8 kilobases), dictated the synthesis of a smaller-sized p53 protein (45 kilodaltons). The p53 proteins synthesized in vitro immunoprecipitated efficiently with human-specific anti-p53 antibodies. Genomic analysis of human DNA revealed the presence of a single p53 gene residing within two EcoRI fragments. Heteroduplex analysis between the full-length cDNA clone p53-H7 and the cloned p53 gene indicated the presence of seven major exons.

scholarly journals A highly conserved mouse gene with a propensity to form pseudogenes in mammals.

1988 ◽  
Vol 8 (7) ◽  
pp. 2797-2803 ◽  
Author(s):  
D L Heller ◽  
K M Gianola ◽  
L A Leinwand
Keyword(s):  
Dna Sequence ◽  
Cdna Clone ◽  
Restriction Analysis ◽  
In Vitro Transcription ◽  
Open Reading Frame ◽  
Cdna Clones ◽  
Reading Frame ◽  
Mouse Cdna ◽  
Processed Pseudogenes

A mouse cDNA clone corresponding to an abundantly transcribed poly(A)+ mRNA was found to be represented by 200 copies in mammalian genomes. To understand the origin and nature of this sequence family, we studied two genomic members and two cDNA clones from mouse liver. The DNA sequence of the coding strand of a full-length cDNA clone was shown to have an open reading frame capable of encoding a 25-kilodalton polypeptide that has not been previously described. In vitro transcription-translation experiments verified the presence of an open reading frame encoding a protein of the predicted size. Restriction analysis of genomic DNA and DNA sequence analysis of genomic clones indicated that many of the 200 members of this family represent processed pseudogenes, with one or a small number of active structural genes. The vast majority of the genomic copies are heterogeneous in length, truncated at their 5' ends with respect to the mRNA, and do not appear to have intervening sequences. Two distinct genomic members of this family were sequenced and found to represent incomplete copies of the mRNA. Both are 5' truncated at slightly different points with respect to the mRNA. Both pseudogenes have multiple base changes, insertions, and deletions relative to the mRNA, and one of them encodes the poly(A) tail of the mRNA. The expression of this gene family is highest in rapidly dividing cells such as early mouse embryos and testis, but was seen in all tissues tested. This gene shows extremely high sequence conservation, extending to chicken, amphibian, and nematode genomes. Surprisingly, the gene appears to exist in only one copy in these organisms.

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

Infectivity of in vitro transcripts of Johnsongrass mosaic potyvirus full-length cDNA clones in maize and sorghum

2003 ◽  
Vol 148 (3) ◽  
pp. 563-574 ◽  
Author(s):  
K.-S. Kim ◽  
H.-Y. Oh ◽  
S. Suranto ◽  
E. Nurhayati ◽  
K. H. Gough ◽  
...  
Keyword(s):  
Full Length ◽  
Cdna Clones ◽  
Full Length Cdna ◽  
In Vitro Transcripts
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