5′-flanking sequences that inhibit in vitro transcription of a Xenopus laevis tRNA gene

Cell ◽  
1983 ◽  
Vol 34 (3) ◽  
pp. 881-890 ◽  
Author(s):  
Robert A. Hipskind ◽  
Stuart G. Clarkson
Keyword(s):  
Xenopus Laevis ◽  
Trna Gene ◽  
In Vitro Transcription ◽  
Flanking Sequences

scholarly journals Transcriptionally inactive oocyte-type 5S RNA genes of Xenopus laevis are complexed with TFIIIA in vitro.

1987 ◽  
Vol 7 (10) ◽  
pp. 3503-3510 ◽  
Author(s):  
L J Peck ◽  
L Millstein ◽  
P Eversole-Cire ◽  
J M Gottesfeld ◽  
A Varshavsky
Keyword(s):  
Xenopus Laevis ◽  
Differential Expression ◽  
Gene Transcription ◽  
Nuclear Extract ◽  
In Vitro Transcription ◽  
5S Rna ◽  
Differential Binding ◽  
5S Rna Genes ◽  
Rna Genes

An extract from whole oocytes of Xenopus laevis was shown to transcribe somatic-type 5S RNA genes approximately 100-fold more efficiently than oocyte-type 5S RNA genes. This preference was at least 10-fold greater than the preference seen upon microinjection of 5S RNA genes into oocyte nuclei or upon in vitro transcription in an oocyte nuclear extract. The approximately 100-fold transcriptional bias in favor of the somatic-type 5S RNA genes observed in vitro in the whole oocyte extract was similar to the transcriptional bias observed in developing Xenopus embryos. We also showed that in the whole oocyte extract, a promoter-binding protein required for 5S RNA gene transcription, TFIIIA, was bound both to the actively transcribed somatic-type 5S RNA gene and to the largely inactive oocyte-type 5S RNA genes. These findings suggest that the mechanism for the differential expression of 5S RNA genes during Xenopus development does not involve differential binding of TFIIIA to 5S RNA genes.

Structure and in vitro transcription of tRNA gene clusters containing the primers of MuLV reverse transcriptase

1986 ◽  
Vol 158 (3) ◽  
pp. 437-442 ◽  
Author(s):  
Tommaso RUSSO ◽  
Francesco COSTANZO ◽  
Adriana OLIVA ◽  
Rosario AMMENDOLA ◽  
Angela DUILIO ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Trna Gene ◽  
Gene Clusters ◽  
In Vitro Transcription

scholarly journals Structure and in vitro transcription of a glycine tRNA gene from Bombyx mori.

1984 ◽  
Vol 3 (7) ◽  
pp. 1547-1552 ◽  
Author(s):  
A. Fournier ◽  
M.A. Guérin ◽  
J. Corlet ◽  
S.G. Clarkson
Keyword(s):  
Bombyx Mori ◽  
Trna Gene ◽  
In Vitro Transcription

scholarly journals A DNA-binding protein is required for termination of transcription by RNA polymerase I in Xenopus laevis.

1990 ◽  
Vol 10 (6) ◽  
pp. 2793-2800 ◽  
Author(s):  
B McStay ◽  
R H Reeder
Keyword(s):  
Dna Binding ◽  
Xenopus Laevis ◽  
Rna Polymerase ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
In Vitro Transcription ◽  
Rna Polymerase I ◽  
Polymerase I ◽  
Terminator Sequence

We describe a partially fractionated in vitro transcription system from Xenopus laevis for the assay of transcription termination by RNA polymerase I. Termination in vitro was found to require a specific terminator sequence in the DNA and a DNA-binding protein fraction that produces a footprint over the terminator sequence.

scholarly journals Transcriptionally inactive oocyte-type 5S RNA genes of Xenopus laevis are complexed with TFIIIA in vitro

1987 ◽  
Vol 7 (10) ◽  
pp. 3503-3510
Author(s):  
L J Peck ◽  
L Millstein ◽  
P Eversole-Cire ◽  
J M Gottesfeld ◽  
A Varshavsky
Keyword(s):  
Xenopus Laevis ◽  
Differential Expression ◽  
Gene Transcription ◽  
Nuclear Extract ◽  
In Vitro Transcription ◽  
5S Rna ◽  
Differential Binding ◽  
5S Rna Genes ◽  
Rna Genes

An extract from whole oocytes of Xenopus laevis was shown to transcribe somatic-type 5S RNA genes approximately 100-fold more efficiently than oocyte-type 5S RNA genes. This preference was at least 10-fold greater than the preference seen upon microinjection of 5S RNA genes into oocyte nuclei or upon in vitro transcription in an oocyte nuclear extract. The approximately 100-fold transcriptional bias in favor of the somatic-type 5S RNA genes observed in vitro in the whole oocyte extract was similar to the transcriptional bias observed in developing Xenopus embryos. We also showed that in the whole oocyte extract, a promoter-binding protein required for 5S RNA gene transcription, TFIIIA, was bound both to the actively transcribed somatic-type 5S RNA gene and to the largely inactive oocyte-type 5S RNA genes. These findings suggest that the mechanism for the differential expression of 5S RNA genes during Xenopus development does not involve differential binding of TFIIIA to 5S RNA genes.

Accurate in vitro transcription of Xenopus laevis mitochondrial DNA from two bidirectional promoters

1986 ◽  
Vol 6 (7) ◽  
pp. 2543-2550
Author(s):  
D F Bogenhagen ◽  
B K Yoza
Keyword(s):  
Mitochondrial Dna ◽  
Xenopus Laevis ◽  
Rna Polymerase ◽  
Consensus Sequence ◽  
In Vitro Transcription ◽  
Xenopus Laevis Oocytes ◽  
Mitochondrial Rna ◽  
Bidirectional Promoters

The mitochondrial RNA polymerase from Xenopus laevis oocytes was partially purified by heparin-Sepharose chromatography and phosphocellulose chromatography. This RNA polymerase preparation specifically initiated the transcription of X. laevis mitochondrial DNA (mtDNA) from two bidirectional promoters contained within a 123-base-pair segment of the mtDNA between the heavy-strand replication origin and the rRNA cistrons. Transcription in vitro initiated from precisely the same start sites previously mapped as initiation sites for transcription in vivo. At each of the four sites, initiation occurred within a conserved nucleotide sequence, ACPuTTATA. This consensus sequence is not related to promoters for transcription of human mtDNA.

scholarly journals Molecular characterization of chicken syndecan-2 proteoglycan

2002 ◽  
Vol 366 (2) ◽  
pp. 481-490 ◽  
Author(s):  
Ligong CHEN ◽  
John R. COUCHMAN ◽  
Jacqueline SMITH ◽  
Anne WOODS
Keyword(s):  
Xenopus Laevis ◽  
Transmembrane Domain ◽  
Heparan Sulphate ◽  
Fibroblast Cell ◽  
In Vitro Transcription ◽  
Northern Analysis ◽  
Nucleotide Polymorphisms ◽  
Chicken Embryonic Fibroblast ◽  
Embryonic Fibroblast

A partial syndecan-2 sequence (147bp) was obtained from chicken embryonic fibroblast poly(A)+ RNA by reverse transcription–PCR. This partial sequence was used to produce a 5′-end-labelled probe. A chicken liver cDNA library was screened with this probe, and overlapping clones were obtained encompassing the entire cDNA of 3kb. The open reading frame encodes a protein of 201 amino acids. The cytoplasmic domain is identical with that of mammalian syndecan-2, and highly similar to those of Xenopus laevis and zebrafish syndecan-2. The transmembrane domain is identical with that of mammalian and zebrafish syndecan-2, and highly similar to that of Xenopus laevis syndecan-2. The ectodomain is 45–62% identical with that of zebrafish, Xenopus laevis and mammalian syndecan-2. Two coding single nucleotide polymorphisms were observed. In vitro transcription and translation yielded a product of 30kDa. Western blotting of chicken embryonic fibroblast cell lysates with species-specific monoclonal antibody mAb 8.1 showed that chicken syndecan-2 is substituted with heparan sulphate, and that the major form of chicken syndecan-2 isolated from chicken fibroblasts is consistent with the formation of SDS-resistant dimers, which is common for syndecans. A 5′-end-labelled probe hybridized to two mRNA species in chicken embryonic fibroblasts, while Northern analysis with poly(A)+ RNAs from different tissues of chicken embryos showed wide and distinct distributions of chicken syndecan-2 during embryonic development. This pattern was different from that reported for syndecan-4, but consistent with the roles of syndecan-2 in neural maturation and in mesenchymal–matrix interactions.

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