Characterisation of Two cDNAs Encoding Carbonic Anhydrase in Maize Leaves

1997 ◽  
Vol 24 (4) ◽  
pp. 451 ◽  
Author(s):  
James N. Burnell ◽  
Martha Ludwig
Keyword(s):  
Carbonic Anhydrase ◽  
Leader Sequence ◽  
Open Reading Frame ◽  
Coding Region ◽  
Reading Frame ◽  
Maize Leaf ◽  
Repeat Sequences ◽  
Maize Leaves ◽  
Molecular Masses ◽  
Triton X

Two distinct cDNAs encoding carbonic anhydrase (CA) were isolated from a maize λgt 11 library. One of these cDNAs, CA1, which consists of a 5´-leader sequence, three repeat sequences and a 3´-non-coding region, is predicted to encode an open reading frame for a polypeptide of 71.3 kDa. The other cDNA, CA2, which has a 5´-leader sequence containing a 276 bp insert compared to CA1, two repeat sequences and a 3´-non-coding region, is predicted to encode an open reading frame for a polypeptide of 59.2 kDa. Nucleotide sequence alignment analysis indicates that the two repeat sequences of CA2 are homologous with repeat sequences 1 and 3 of CA1, respectively. Four protein bands, with apparent molecular masses of 52, 47, 28 and 27 kDa, were evident in western blot analyses of crude extracts of maize leaf tissue prepared in the absence of Triton X-100 and two protein bands, with apparent molecular masses of 27 and 28 kDa, were detected in western blots of crude extracts prepared in the presence of Triton X-100. These results indicate either that both CA1 and CA2 mRNAs are only partially translated, that the CA1 and CA2 proteins are processed, or a combination of both of these alternatives. Two maize leaf CA1 and CA2 mRNAs detected on northern blots are longer than any other plant CA mRNA reported to date. Possible roles for the two CA isozymes in maize leaves are discussed.

Cloning of a gar (Lepisosteus osseus) GH cDNA: trends in actinopterygian GH structure

1996 ◽  
Vol 16 (1) ◽  
pp. 73-80 ◽  
Author(s):  
D A Rubin ◽  
J H Youson ◽  
L E Marra ◽  
R M Dores
Keyword(s):  
Cdna Library ◽  
Untranslated Region ◽  
Signal Sequence ◽  
Leader Sequence ◽  
Open Reading Frame ◽  
Russian Sturgeon ◽  
Reading Frame ◽  
Terminal Codon

ABSTRACT A cDNA containing the sequence of GH was cloned and sequenced from a pituitary cDNA library for the holostean fish Lepisosteus osseus (common name: gar). The gar GH cDNA contained an open reading frame of 633 nucleotides and a 3′ untranslated region (including the terminal codon TAG) of 1058 nucleotides. The overall length of the gar GH cDNA including leader sequence, signal sequence, hormone sequence and 3′ untranslated region was 1713 nucleotides. Thus, the gar GH cDNA is the largest vertebrate GH cDNA yet cloned. A comparison of GH sequences from ancient (holostean fishes — gar and bowfin; one chondrostean fish — the Russian sturgeon) and more modern (27 species of teleosts) members of class Actinopterygii indicate that members of this class have maintained many of the invariant residues deemed necessary for GH folding motifs (intramolecular relationships) observed in mammals.

Pax-6, a murine paired box gene, is expressed in the developing CNS

Development ◽  
1991 ◽  
Vol 113 (4) ◽  
pp. 1435-1449 ◽  
Author(s):  
C. Walther ◽  
P. Gruss
Keyword(s):  
Amino Acids ◽  
Neural Tube ◽  
Multigene Family ◽  
Ventricular Zone ◽  
Open Reading Frame ◽  
Cdna Clones ◽  
Coding Region ◽  
Paired Domain ◽  
Reading Frame ◽  
Pax Genes

A multigene family of paired-box-containing genes (Pax genes) has been identified in the mouse. In this report, we describe the expression pattern of Pax-6 during embryogenesis and the isolation of cDNA clones spanning the entire coding region. The Pax-6 protein consists of 422 amino acids as deduced from the longest open reading frame and contains, in addition to the paired domain, a paired-type homeodomain. Beginning with day 8 of gestation, Pax-6 is expressed in discrete regions of the forebrain and the hindbrain. In the neural tube, expression is mainly confined to mitotic active cells in the ventral ventricular zone along the entire anteroposterior axis starting at day 8.5 of development. Pax-6 is also expressed in the developing eye, the pituitary and the nasal epithelium.

Retroviral insertions in the murine His-1 locus activate the expression of a novel RNA that lacks an extensive open reading frame

1994 ◽  
Vol 14 (3) ◽  
pp. 1743-1751
Author(s):  
D S Askew ◽  
J Li ◽  
J N Ihle
Keyword(s):  
Cell Lines ◽  
Wild Mouse ◽  
Myeloid Cell ◽  
Single Copy ◽  
Open Reading Frame ◽  
Chromosome 2 ◽  
Coding Region ◽  
Reading Frame ◽  
Novel Gene ◽  
Myeloid Leukemias

The His-1 locus is a common site of viral insertion in murine myeloid leukemias induced by the wild mouse ecotropic retrovirus, CasBrM. In this report, we describe the cloning of a novel gene at the His-1 locus and show that His-1 expression is associated with the transformed phenotype. Northern (RNA) blot analysis identified His-1 transcripts in four transformed myeloid cell lines but in no normal tissues examined. Two of these cell lines were derived from retrovirus-induced myeloid leukemias that harbor integrated proviruses which drive His-1 gene expression by promoter insertion. The two other cell lines expressed a discrete 3-kb His-1 RNA that is derived from a novel gene consisting of three exons that span 6 kb on mouse chromosome 2. The His-1 gene is conserved as a single-copy sequence in multiple vertebrate species and is expressed as a spliced and polyadenylated RNA. A protein-coding region is not evident from analysis of the His-1 sequence because of the presence of multiple small open reading frames, none of which are greater than 219 bp. This lack of an extensive open reading frame is an unusual feature that is shared by other RNA molecules believed to function in the absence of translation.

The Saccharomyces cerevisiae genome contains functional and nonfunctional copies of transposon Ty1

1988 ◽  
Vol 8 (4) ◽  
pp. 1432-1442 ◽  
Author(s):  
J D Boeke ◽  
D Eichinger ◽  
D Castrillon ◽  
G R Fink
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Long Terminal Repeat ◽  
Missense Mutation ◽  
Dna Sequence ◽  
Open Reading Frame ◽  
Yeast Genome ◽  
Reading Frame ◽  
Ty Elements ◽  
Ty Element ◽  
Repeat Sequences

Saccharomyces cerevisiae Ty elements are transposons closely related to retroviruses. The DNA sequence of a functional Ty element (TyH3) is presented. The long terminal repeat sequences are different, suggesting that TyH3 is a recombinant Ty element. A chromosomal Ty element near the LYS2 gene, Ty173, was found to be nonfunctional, even though it has no detectable insertions or deletions. The defect in Ty173 transposition is caused by a missense mutation giving rise to a Leu-to-Ile substitution in the TYB (pol) open reading frame. Several chromosomal Ty elements carry this lesion in their DNA, indicating that nonfunctional Ty elements are common in the yeast genome.

scholarly journals Mapping of Transcription Termination within the S Segment of SFTS Phlebovirus Facilitated Generation of NSs Deletant Viruses

2017 ◽  
Vol 91 (16) ◽  
Author(s):  
Benjamin Brennan ◽  
Veronica V. Rezelj ◽  
Richard M. Elliott
Keyword(s):  
Virus Infection ◽  
Reverse Genetics ◽  
Fluorescent Protein ◽  
Nonstructural Protein ◽  
Transcription Termination ◽  
Severe Disease ◽  
Open Reading Frame ◽  
Coding Region ◽  
Reading Frame ◽  
Green Fluorescent

ABSTRACT SFTS phlebovirus (SFTSV) is an emerging tick-borne bunyavirus that was first reported in China in 2009. Here we report the generation of a recombinant SFTSV (rHB29NSsKO) that cannot express the viral nonstructural protein (NSs) upon infection of cells in culture. We show that rHB29NSsKO replication kinetics are greater in interferon (IFN)-incompetent cells and that the virus is unable to suppress IFN induced in response to viral replication. The data confirm for the first time in the context of virus infection that NSs acts as a virally encoded IFN antagonist and that NSs is dispensable for virus replication. Using 3′ rapid amplification of cDNA ends (RACE), we mapped the 3′ end of the N and NSs mRNAs, showing that the mRNAs terminate within the coding region of the opposite open reading frame. We show that the 3′ end of the N mRNA terminates upstream of a 5′-GCCAGCC-3′ motif present in the viral genomic RNA. With this knowledge, and using virus-like particles, we could demonstrate that the last 36 nucleotides of the NSs open reading frame (ORF) were needed to ensure the efficient termination of the N mRNA and were required for recombinant virus rescue. We demonstrate that it is possible to recover viruses lacking NSs (expressing just a 12-amino-acid NSs peptide or encoding enhanced green fluorescent protein [eGFP]) or an NSs-eGFP fusion protein in the NSs locus. This opens the possibility for further studies of NSs and potentially the design of attenuated viruses for vaccination studies. IMPORTANCE SFTS phlebovirus (SFTSV) and related tick-borne viruses have emerged globally since 2009. SFTSV has been shown to cause severe disease in humans. For bunyaviruses, it has been well documented that the nonstructural protein (NSs) enables the virus to counteract the human innate antiviral defenses and that NSs is one of the major determinants of virulence in infection. Therefore, the use of reverse genetics systems to engineer viruses lacking NSs is an attractive strategy to rationally attenuate bunyaviruses. Here we report the generation of several recombinant SFTS viruses that cannot express the NSs protein or have the NSs open reading frame replaced with a reporter gene. These viruses cannot antagonize the mammalian interferon (IFN) response mounted to virus infection. The generation of NSs-lacking viruses was achieved by mapping the transcriptional termination of two S-segment-derived subgenomic mRNAs, which revealed that transcription termination occurs upstream of a 5′-GCCAGCC-3′ motif present in the virus genomic S RNA.

scholarly journals Nucleotide sequences of segments 1, 3 and 4 of the genome of Bombyx mori cypovirus 1 encoding putative capsid proteins VP1, VP3 and VP4, respectively

2002 ◽  
Vol 83 (6) ◽  
pp. 1477-1482 ◽  
Author(s):  
Kyoji Hagiwara ◽  
Shujing Rao ◽  
Simon W. Scott ◽  
Gerald R. Carner
Keyword(s):  
Amino Acids ◽  
Bombyx Mori ◽  
Nucleotide Sequences ◽  
Open Reading Frame ◽  
Capsid Proteins ◽  
Structural Proteins ◽  
Reading Frame ◽  
The Family ◽  
Molecular Masses ◽  
Rice Ragged Stunt Virus

The complete nucleotide sequences of genomic segments S1, S3 and S4 from Bombyx mori cypovirus 1 (BmCPV-1) have been determined. The segments consisted of 4190, 3846 and 3262 nucleotides encoding putative proteins of 1333, 1239 and 1058 amino acids with molecular masses of approximately 148, 140 and 120 kDa (p148, p140 and p120, respectively). All segments possess a single open reading frame. Homology searches showed that all three proteins have homologies to proteins of Rice ragged stunt virus, a member of the genus Oryzavirus within the family Reoviridae. Partial homologies of p140 to structural proteins in other viruses were also found. The predicted molecular masses and the homologies with structural proteins in other viruses lead us to suggest that S1, S3 and S4 encode the capsid proteins VP1, VP3, and VP4, respectively, of BmCPV-1.

scholarly journals Comparative Biology of Two Natural Variants of the IncQ-2 Family Plasmids, pRAS3.1 and pRAS3.2

2009 ◽  
Vol 191 (20) ◽  
pp. 6436-6446 ◽  
Author(s):  
Wesley Loftie-Eaton ◽  
Douglas E. Rawlings
Keyword(s):  
Copy Number ◽  
Promoter Region ◽  
Plasmid Stability ◽  
Inducible Promoter ◽  
Open Reading Frame ◽  
Comparative Biology ◽  
Incompatibility Group ◽  
Reading Frame ◽  
Repeat Sequences ◽  
Natural Variants

ABSTRACT Plasmids pRAS3.1 and pRAS3.2 are two closely related, natural variants of the IncQ-2 plasmid family that have identical plasmid backbones except for two differences. Plasmid pRAS3.1 has five 6-bp repeat sequences in the promoter region of the mobB gene and four 22-bp iterons in its oriV region, whereas pRAS3.2 has only four 6-bp repeats and three 22-bp iterons. Plasmid pRAS3.1 was found to have a higher copy number than pRAS3.2, and we show that the extra 6-bp repeat results in an increase in mobB and downstream mobA/repB expression. Placement of repB (primase) behind an arabinose-inducible promoter in trans resulted in an increase in repB expression and an approximately twofold increase in the copy number of plasmids with identical numbers of 22-bp iterons. The pRAS3 plasmids were shown to have a previously unrecognized toxin-antitoxin plasmid stability module within their replicons. The ability of the pRAS3 plasmids to mobilize the oriT regions of two other plasmids of the IncQ-2 family, pTF-FC2 and pTC-F14, suggested that the mobilization proteins pRAS3 are relaxed and can mobilize oriT regions with substantially different sequences. Plasmids pRAS3.1 and pRAS3.2 were highly incompatible with plasmids pTF-FC2 and pTC-F14, and this incompatibility was removed on inactivation of an open reading frame situated downstream of the mobCDE mobilization genes rather than being due to the 22-bp oriV-associated iterons. We propose that the pRAS3 plasmids represent a third, γ incompatibility group within the IncQ-2 family plasmids.

scholarly journals Minimal and Contributing Sequence Determinants of thecis-Acting Locus of Transfer (clt) of Streptomycete Plasmid pIJ101 Occur within an Intrinsically Curved Plasmid Region

2000 ◽  
Vol 182 (23) ◽  
pp. 6834-6841 ◽  
Author(s):  
Matthew J. Ducote ◽  
Shubha Prakash ◽  
Gregg S. Pettis
Keyword(s):  
Transfer Function ◽  
Inverted Repeat ◽  
Regulatory Gene ◽  
Direct Repeat ◽  
Higher Order ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Repeat Sequences ◽  
Inherent Feature ◽  
Close Proximity

ABSTRACT Efficient interbacterial transfer of streptomycete plasmid pIJ101 requires the pIJ101 tra gene, as well as acis-acting plasmid function known as clt. Here we show that the minimal pIJ101 clt locus consists of a sequence no greater than 54 bp in size that includes essential inverted-repeat and direct-repeat sequences and is located in close proximity to the 3′ end of the korB regulatory gene. Evidence that sequences extending beyond the minimal locus and into thekorB open reading frame influence clt transfer function and demonstration that clt-korB sequences are intrinsically curved raise the possibility that higher-order structuring of DNA and protein within this plasmid region may be an inherent feature of efficient pIJ101 transfer.

scholarly journals The Saccharomyces cerevisiae genome contains functional and nonfunctional copies of transposon Ty1.

1988 ◽  
Vol 8 (4) ◽  
pp. 1432-1442 ◽  
Author(s):  
J D Boeke ◽  
D Eichinger ◽  
D Castrillon ◽  
G R Fink
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Long Terminal Repeat ◽  
Missense Mutation ◽  
Dna Sequence ◽  
Open Reading Frame ◽  
Yeast Genome ◽  
Reading Frame ◽  
Ty Elements ◽  
Ty Element ◽  
Repeat Sequences

Saccharomyces cerevisiae Ty elements are transposons closely related to retroviruses. The DNA sequence of a functional Ty element (TyH3) is presented. The long terminal repeat sequences are different, suggesting that TyH3 is a recombinant Ty element. A chromosomal Ty element near the LYS2 gene, Ty173, was found to be nonfunctional, even though it has no detectable insertions or deletions. The defect in Ty173 transposition is caused by a missense mutation giving rise to a Leu-to-Ile substitution in the TYB (pol) open reading frame. Several chromosomal Ty elements carry this lesion in their DNA, indicating that nonfunctional Ty elements are common in the yeast genome.

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