scholarly journals Antisense Orientation

2020 ◽  
Author(s):  
Keyword(s):  
Antisense Orientation

scholarly journals Altered lignin composition in transgenic tobacco expressing O-methyltransferase sequences in sense and antisense orientation

1995 ◽  
Vol 8 (4) ◽  
pp. 465-477 ◽  
Author(s):  
Rossitza Atanassova ◽  
Noelle Favet ◽  
Francoise Martz ◽  
Brigitte Chabbert ◽  
Marie-Therese Tollier ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Antisense Orientation ◽  
Lignin Composition

An antisense promoter of the murine c-myc gene is localized within intron 2

1992 ◽  
Vol 12 (3) ◽  
pp. 1324-1329
Author(s):  
D B Spicer ◽  
G E Sonenshein
Keyword(s):  
Antisense Transcription ◽  
In Vitro Transcription ◽  
Region Gene ◽  
S1 Nuclease ◽  
Gene Translocation ◽  
Antisense Orientation ◽  
Nuclear Extracts ◽  
Myc Gene ◽  
Intron 2

Previously we have demonstrated the existence of stable transcripts from the noncoding strand of a rearranged c-myc gene in murine plasmacytomas in which the oncogene has translocated to an immunoglobulin constant-region gene element (M. Dean, R. B. Kent, and G. E. Sonenshein, Nature [London] 305:443-446, 1983). The resulting RNAs are chimeric, containing c-myc antisense and immunoglobulin sense sequences. A normal unrearranged murine c-myc gene is transcribed in the antisense orientation throughout much of the gene; however, stable transcripts have not been detected. In this study, using Northern (RNA) blot, S1 nuclease, and primer extension analyses, we have mapped the 5' end of the stable chimeric transcripts to a site 175 bp from the start of exon 3, within intron 2 of the c-myc gene. In vitro transcription assays with constructs containing this site and 400 bp upstream, in the antisense orientation, and nuclear extracts from plasmacytoma cells, as well as a number of cell lines with normal unrearranged c-myc genes, indicated that this promoter was functional. This finding was confirmed in transient transfection assays using the antisense promoter linked to the chloramphenicol acetyltransferase reporter gene. These results suggest that a normal promoter of antisense transcription is used following c-myc gene translocation.

scholarly journals Mechanism(s) of butyrate transport in Caco-2 cells: role of monocarboxylate transporter 1

2000 ◽  
Vol 279 (4) ◽  
pp. G775-G780 ◽  
Author(s):  
Christos Hadjiagapiou ◽  
Larry Schmidt ◽  
Pradeep K. Dudeja ◽  
Thomas J. Layden ◽  
Krishnamurthy Ramaswamy
Keyword(s):  
Short Chain Fatty Acid ◽  
Rnase Protection Assay ◽  
Significant Inhibition ◽  
Monocarboxylate Transporter ◽  
Rnase Protection ◽  
Monocarboxylate Transporter 1 ◽  
Antisense Orientation ◽  
Transporter Family ◽  
Saturation Kinetics

The short-chain fatty acid butyrate was readily taken up by Caco-2 cells. Transport exhibited saturation kinetics, was enhanced by low extracellular pH, and was Na+independent. Butyrate uptake was unaffected by DIDS; however, α-cyano-4-hydroxycinnamate and the butyrate analogs propionate and l-lactate significantly inhibited uptake. These results suggest that butyrate transport by Caco-2 cells is mediated by a transporter belonging to the monocarboxylate transporter family. We identified five isoforms of this transporter, MCT1, MCT3, MCT4, MCT5, and MCT6, in Caco-2 cells by PCR, and MCT1 was found to be the most abundant isoform by RNase protection assay. Transient transfection of MCT1, in the antisense orientation, resulted in significant inhibition of butyrate uptake. The cells fully recovered from this inhibition by 5 days after transfection. In conclusion, our data showed that the MCT1 transporter may play a major role in the transport of butyrate into Caco-2 cells.

scholarly journals Development of Papaya Breeding Lines with Transgenic Resistance to Papaya ringspot virus

Plant Disease ◽  
2004 ◽  
Vol 88 (4) ◽  
pp. 352-358 ◽  
Author(s):  
Michael J. Davis ◽  
Zhentu Ying
Keyword(s):  
Transgenic Line ◽  
Carica Papaya ◽  
Ringspot Virus ◽  
Papaya Ringspot Virus ◽  
Transgenic Resistance ◽  
Breeding Lines ◽  
Antisense Orientation ◽  
Resistant Lines ◽  
Sense Orientation ◽  
Transgenic Lines

Papaya (Carica papaya) was transformed via Agrobacterium-mediated transformation with four constructs containing either the unmodified or modified coat protein (CP) gene of Florida isolate H1K of Papaya ringspot virus (PRSV). The CP genes were in the sense orientation (S-CP), antisense orientation (AS-CP), sense orientation with a frame-shift mutation (FS-CP), or sense orientation mutated with three-in-frame stop codons (SC-CP). In all, 256 putative transgenic lines with the CP constructs were inoculated mechanically with PRSV H1K. None of the lines was immune to PRSV; however, highly resistant lines were found in each CP transgene group. For breeding purposes, 21 PRSV-resistant lines representing the four transgene constructs were selected and crossed with six papaya genotypes. The lines from the FS-CP and SC-CP transgene groups were highly fertile, but those from the S-CP and AS-CP transgene groups were practically infertile. Plants derived from 54 crosses and representing 17 transgenic lines were planted in the field. After 1 year in the field, 293 of the 1,258 the plants (23.3%) became naturally infected with PRSV; whereas, 29 of 30 of the nontransgenic control plants (96.7%) became infected. The incidence of PRSV infection varied in the R1 progeny depending on both the transgenic line and the nontransgenic parent.

scholarly journals Overexpression of prothymosin α accelerates proliferation and retards differentiation in HL-60 cells

1998 ◽  
Vol 331 (3) ◽  
pp. 753-761 ◽  
Author(s):  
Pilar RODRÍGUEZ ◽  
Juan E. VIÑUELA ◽  
Leoncio ÁLVAREZ-FERNÁNDEZ ◽  
Montserrat BUCETA ◽  
Anxo VIDAL ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Opposite Effect ◽  
Cellular Proliferation ◽  
Nuclear Protein ◽  
Prothymosin Α ◽  
Mammalian Cell Lines ◽  
Antisense Orientation ◽  
Cell Clones ◽  
Proliferation And Differentiation ◽  
Antisense Construct

Prothymosin α (ProTα) is an acidic nuclear protein the expression of which is related to the proliferation and differentiation processes in mammalian cells. In the present study we have stably transfected HL-60 cells, a biological system that allows the study of both proliferation and differentiation, with recombinant vectors encoding sense and antisense ProTα mRNA. In the HL-60 cell clones overexpressing ProTα we observed an acceleration in the growth rate, whereas expression of the antisense orientation showed the opposite effect. Moreover, cell-cycle analysis demonstrated that the G1-phase was shortened in the cells expressing the sense construct. Before studying how ProTα affects differentiation, we showed that the down-regulation of ProTα gene during differentiation occurs in all mammalian cell lines (HL-60, K562, U937, MEL C88, N2A and PC12) analysed. The biological effect evoked by the induction of the ProTα sense vector was the retardation of cell differentiation, although expression of the antisense construct showed no effect on differentiation. In conclusion, our findings provide evidence that ProTα is directly implicated in cellular proliferation and that the maintenance of high levels of ProTα inside HL-60 cells is incompatible with their ability to differentiate.

scholarly journals Lupus antiribosomal P antisera contain antibodies to a small fragment of 28S rRNA located in the proposed ribosomal GTPase center.

1991 ◽  
Vol 174 (3) ◽  
pp. 507-514 ◽  
Author(s):  
J L Chu ◽  
N Brot ◽  
H Weissbach ◽  
K Elkon
Keyword(s):  
28S Rrna ◽  
Direct Role ◽  
Autoantibody Production ◽  
Antisense Orientation ◽  
Form Part ◽  
Rna Fragments ◽  
P Proteins ◽  
Ribosomal P ◽  
Made In

The ribosomal P proteins are necessary for GTPase activity during protein synthesis. In addition to antibodies to the P proteins, sera from lupus patients contain anti-rRNA activity. To determine whether lupus antiribosomal sera recognize the region of 28S rRNA recently proposed to form part of the ribosomal GTPase center, an rRNA fragment corresponding to nucleotides (nt) 1922-2020 was transcribed in vitro and tested for antigenicity. 18 of 24 (75%) lupus sera containing anti-P antibodies, but only 2 of 24 (8%) lupus sera without anti-P, immunoprecipitated this rRNA fragment (p less than 0.001). The binding was specific, since no significant differences were observed between anti-P positive and negative lupus sera in binding to the RNA fragment transcribed in the antisense orientation or to a control region of rRNA. The majority of sera tested protected a rRNA fragment of approximately 68 nucleotides. To evaluate the fine specificity of the anti-28S antibodies, deletions and site-directed mutations were made in the RNA fragment. The anti-28S antisera required nt 1944-1955 for recognition and were remarkably sensitive to destabilizing as well as nondestabilizing mutations in the stems of the RNA fragments. Detection of antiprotein and anti-RNA antibodies directed against a functionally related domain in the ribosome, together with the remarkable specificity of anti-28S antibodies, strongly suggests a direct role for this region of the ribosome in initiating and/or maintaining antiribosomal autoantibody production.

scholarly journals Insertion of an SVA Element in MSH2 as a Novel Cause of Lynch Syndrome

2020 ◽  
Author(s):  
Ciyu Yang ◽  
Yirong Li ◽  
Magan Trottier ◽  
Michael Farrell ◽  
Vikas Rai ◽  
...  
Keyword(s):  
Lynch Syndrome ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Dna Mismatch ◽  
Mmr Genes ◽  
Antisense Orientation ◽  
The Family ◽  
Long Range Pcr ◽  
Early Onset Colorectal Cancer ◽  
Generation Sequencing

Germline mutations in the DNA mismatch repair (MMR) genes cause Lynch syndrome (LS). Insertions of retrotransposons in MMR genes have been reported as a rare cause of LS. Here, we present a novel SINE-VNTR-Alu (SVA) insertion in exon 12 of MSH2 in an individual with early-onset colorectal cancer and strong LS family history. RT-PCR analysis indicated a larger aberrant MSH2 transcript in one of the family members. MSK-IMPACT next-generation sequencing testing and long-range PCR revealed an insertion in MSH2 exon 12 at the c.1972 position in an antisense orientation. The insertion was further characterized as an SVA element approximately 3 kb in length, belonging to the SVA_F1 family of retrotransposons.

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