Chloroplast ribosomal RNA genes in Euglena gracilis exist as three clustered tandem repeats

Rawson, James R.Y.; Kushner, Sidney R.; Vapnek, Daniel; Alton, N. K.; Boerma, Cindy L.

doi:10.1016/0378-1119(78)90032-x

Cited by 76 publications

(28 citation statements)

References 30 publications

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“…Euglena chloroplast rDNA was cloned into the Bam HI restriction endonuclease site of the bacterial plasmid vector pBR313 (20) to yield a recombinant plasmid pVK52 containing a complete chloroplast rDNA repeat 6.2 kbp in length (Fig. 3).…”

Section: Methodsmentioning

confidence: 99%

“…This DNA fragment represents the spacer sequence between the rDNA repeat unit on the chloroplast genome and contains part of the 5S rRNA gene (14,15). An Eco RI chloroplast DNA fragment (Eco RI-P) containing the complete 16S rRNA gene, two tRNA genes, and approximately 6% 23S rRNA was cloned into the vector RSF2124 (14,15,20) to yield the recombinant plasmid pVK49. This Eco RI chloroplast DNA fragment is completely contained in pVK52.…”

Section: Methodsmentioning

confidence: 99%

“…Restriction endonuclease DNA fragments were separated on the basis of mol wt by electrophoresis in agarose slab gels. The gels were prepared in E buffer (36 mm Tris, 30 mm NaH2PO4, and 1 mM EDTA) and run at room temperature at 1.5 to 2.0 v/cm for 12 to 16 h. The gels were stained and photographed as described earlier (20). HindIII A DNA digestion products were used as mol wt markers.…”

Section: Methodsmentioning

confidence: 99%

“…Both the nuclear and chloroplast rDNAs from Euglena gracilis have been cloned and individually isolated as homogenous recombinant DNA molecules (4,20). These cloned rDNAs were used in hybridization studies to demonstrate the presence of specific nucleotide sequence homology between chloroplast and nuclear rDNAs and to localize those regions on restriction endonuclease maps of the two rDNAs.…”

Section: Introductionmentioning

confidence: 99%

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Nucleotide Sequence Homology Exists between the Chloroplast and Nuclear Ribosomal DNAs of Euglena gracilis

Curtis

Rawson²

1982

Plant Physiol.

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The nuclear and chioroplast ribosomal DNAs from Eugkna were shown to have specific regions of nucleotide sequence homology. The regions of homology were identified by hybriization ofrestriction endonuclease DNA fragments of cloned chloroplast and nuclear ribosomal DNAs to one another. The regions of homolo between these two ribosomal DNAs were In that part of the genes that code for the 3' end of the small rRNAs (16S and 19S) and near or at the DNA sequences coding for the 5S RNAs.The nucleotde sequence homology between these regions was estimated to be approximately 94% by the melting point depression of a hybrid formed between the two ribosomal DNAs.Nucleotide sequence homology between chloroplast and nuclear rRNAs has been explored in a variety of plants (1,12,26,28). All of these studies have shown hybridization between the cytoplasmic rRNA and chloroplast DNA or reciprocally between the chloroplast rRNA and the nuclear DNA. These results have been interpreted in three ways: (a) nucleotide sequence homology exists between the nuclear and chloroplast rDNAs2; (b) the cytoplasmic and/or chloroplast rRNAs used for these studies were contaminated with one another, or (c) the chloroplast and nuclear DNAs to which the different rRNAs were hybridized were contaminated with one another.Final resolution of the question of nucleotide sequence homology between nuclear and chloroplast rDNAs required the use of absolutely pure preparations of nuclear and chloroplast rDNAs. Both the nuclear and chloroplast rDNAs from Euglena gracilis have been cloned and individually isolated as homogenous recombinant DNA molecules (4,20). These cloned rDNAs were used in hybridization studies to demonstrate the presence of specific nucleotide sequence homology between chloroplast and nuclear rDNAs and to localize those regions on restriction endonuclease maps of the two rDNAs. MATERIALS AND METHODSConstruction of Recombinant Plasmids. Euglena chloroplast rDNA was cloned into the Bam HI restriction endonuclease site of the bacterial plasmid vector pBR313 (20) to yield a recombinant plasmid pVK52 containing a complete chloroplast rDNA repeat 6.2 kbp in length (Fig. 3). This plasmid was used to construct the plasmid pECRBl which contains a 0.85-kbp nucleotide sequence

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Nucleotide Sequence Homology Exists between the Chloroplast and Nuclear Ribosomal DNAs of Euglena gracilis

Curtis

Rawson²

1982

Plant Physiol.

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“…Although the extent of transciption of protein genes could be in the range of 50 000 base pairs [1,2], no protein genes have been mapped on the circular genome so far. On the other hand, all rRNA genes [3][4][5][6] and several of the tRNA genes [5][6][7][8] have been mapped.…”

Section: Introduction 2 Materials and Methodsmentioning

confidence: 99%

Analysis of Euglena gracilis chloroplast DNA

et al. 1981

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Selective in vitro transcription of chloroplast genes

Gruissem

Narita

Greenberg

et al. 1983

J of Cellular Biochemistry

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Transcription of Euglena gracilis chloroplast genes has been investigated by using in vitro transcription systems. A DNA-dependent RNA polymerase responsible for the transcription of rRNA genes has been isolated as a nucleoprotein complex (transcriptionally active chromosome). The RNA polymerase remains tightly bound to the chloroplast DNA template and does not initiate transcription with cloned chloroplast genes. A transcriptionally active extract has been prepared from intact Euglena chloroplasts. The soluble RNA polymerase in this extract recognizes cloned chloroplast tRNA genes and tRNA-sized products have been detected after transcription. The tRNA-sized molecules specifically hybridize to the tRNA genes in the plasmid DNA. At least five tRNA-sized products have been identified from transcription of a trnY1trnH1-trnM1-trnE1-trnW1-trnG1 cluster. Evidence is also presented that processing enzymes in the chloroplast-extract can recognize a polycistronic tRNAVal-tRNAAsn-tRNAArg precursor and process it into tRNA-sized molecules. Truncated templates have been used to demonstrate that the chloroplast tRNA genes are actively transcribed. From a comparison of 5' flanking sequences in chloroplast tRNA genes, a consensus sequence which might function as a promoter, has been identified. The properties of the RNA polymerase involved in the transcription of chloroplast rRNA genes and tRNA genes have been investigated and compared.

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Chloroplast ribosomal RNA genes in Euglena gracilis exist as three clustered tandem repeats

Cited by 76 publications

References 30 publications

Nucleotide Sequence Homology Exists between the Chloroplast and Nuclear Ribosomal DNAs of Euglena gracilis

Nucleotide Sequence Homology Exists between the Chloroplast and Nuclear Ribosomal DNAs of Euglena gracilis

Analysis of Euglena gracilis chloroplast DNA

Selective in vitro transcription of chloroplast genes

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