Organization and Expression of Plastid Genomes

Bohnert, Hans J.; Crouse, Edwin J.; Schmitt, Jürgen

doi:10.1007/978-3-642-68347-3_14

Cited by 39 publications

(29 citation statements)

References 241 publications

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“…to 38% (6), and that this also applies to the individual fragments isolated from barley cpDNA the hybridizations were performed at approximately Tm-21 *C. The subsequent stringent washings of the filters were performed in 0.015 M-NaC1, 1.5 mM-Na-citrate, 0.2% (w/v) Nadodecyl sulphate at 55 *C, corresponding to T m-12 ~ Results of the hybridizations performed with radioactive probes, made from the fragments described in section 3. Figure 8, the data suggest that hybridization conditions may have been too stringent in some cases (b, c, d) and maybe not overly stringent in others (a, e, f, k etc.).…”

Section: Probe Plasmid Hindlii Psti Pvullmentioning

confidence: 99%

“…This suggests that the Pstl site separating the t-and the s-fragments is very close to the 23S rRNA fragmentation point. Furthermore, since the s-fragment subclone pHvC68 gives a hybridization response quite similar to the s-fragment itself, it is possible that the three 3.1-3.5 kb transcripts are different 23S rRNA precursors, which may also include a 4.5S rR_NA (6).…”

Section: The Inverted Repeat and The Small Singlementioning

confidence: 99%

“…Fragment pattern analyses with restriction endonucleases have revealed that chloroplast DNAs (cpDNA) are circular, consist of 124,000 to 180,000 basepairs and frequently contain an inverted repeat of 20,000 to 25,000 basepairs. DNA-RNA hybridization studies showed the inverted repeat to house the genes for ribosomal RNAs (23S, 16S, 5S and others), the rrn-genes (6). One exception to this general organization is Euglena gracilis, which contains a triple tandem repeat of the rRNA cistrons and an extra copy of the 16S rRNA gene.…”

Section: Introducrlonmentioning

confidence: 99%

“…The cloned fragments were employed both in coupled transcriptiontranslation assays and for hybrid-selection of mRNAs followed by in vitro translations to identify the structural genes for the following chloroplast polypeptides: The large subunit of ribulosebisphosphate carboxylase (rbcL) (32,54), ATP synthetase CF~ subunit ct (atpA) (50), subunit [3 and subunit 8 (atpB/atpE) (27,53), cytochrome f (pet) and the subunitIII of ATP synthetase CF 0 (atpH) (21), and one light inducible 32,000 dalton thylakoid protein (psbA) (6). The nucleotide sequences for several of these genes have been or are currently being established (42,55).…”

Section: Introducrlonmentioning

confidence: 99%

“…In vitro hybridizations between chloroplast RNA and chloroplast DNA has indicated that 40-60% of the chloroplast genome is expressed as transcripts (6,33). This would correspond to transcription of about one strand of the genome.…”

Section: Introducrlonmentioning

confidence: 99%

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The barley chloroplast genome: Physical structure and transcriptional activity in vivo

Poulsen

1983

Carlsberg Res. Commun.

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Keywords: Chloroplast DNA, restriction endonucleases, the large subunit of ribulose bisphosphate carboxylase, chloroplast RNA, formaldehyde agarose gels, Northern blotting, filter hybridization, light induced transcription.DNA has been isolated from barley chloroplasts and analyzed with restriction endonucleases PstI, Sail, PvulI and HindIII. Fragments obtained by digestion with PstI and HindIII have been inserted in the bacterial transformation vectors pBR325 and pBR322 and amplified in Escherichia coli. The inserts of the recombinant plasmids have been mapped with the same restriction enzymes as the intact chloroplast DNA yielding a physical map for the barley chloroplast genome. It is a circular molecule about 133,000 basepairs in size, which is equivalent to the chloroplast genomes of other gramineae such as wheat and maize. Like these it has an inverted repeat, of about 20,900 basepairs, containing the genes for the ribosomal RNAs. There are many similarities of the barley, wheat and maize genomes with respect to recognition sites for the enzymes PstI, SalI and PvuII. Heterologous hybridization with a probe containing parts of the genes for the large subunit of ribulose bisphosphate carboxylase and for the dicistronic mRNA for two ATPsynthetase CF~ subunits, reveal that the position and organization of these genes in the barley chloroplast DNA are the same as found for maize and wheat.Selected chloroplast DNA fragments isolated from the recombinant plasmids and covering about 80% of the genome have been used for hybridization to RNA purified from plastids of dark grown and 8 hours illuminated seedlings. The RNA was electrophoretically separated in denaturing agarose gels and subsequently Northern blotted onto DBM-paper. The DNA fragments were nick translated and hybridized to the filterbound RNA. A total of 70 transcripts ranging in size from 350 nucleotides to more than 6,000 were identified. Of these 11 were synthesized only after illumination of the seedlings. The transcripts encoding the large subunit of ribulose bisphosphate carboxylase and the ATP synthetase CFt subunits [3 and e have been tentatively identified. About 30 transcripts are larger than 3,000 nucleotides, indicating either the existence of a large number of polycistronic mRNAs or various forms of precursor RNAs.Abbreviations: bp = basepairs; DBM-= diazobenzyloxymethyl-; DCCD = dicyclohexylcarbodiimide; DCMU = dichlorophenyl-dimethyl-urea; kb = kilobases; kbp = ldlobasepairs; LS = large subunit; NBM-= nitrobenzyloxymethyl-; NBPC = nitrobenzyloxy-methylpyridinium chloride;pHvC = recombinant plasmids carrying Hordeum vulgare chloroplast DNA fragments; RuBPCase = ribulose bisphosphate carboxylase. Genes: rbcL = the gene for RuBPCase LS; atpA = the gene for ATP synthetase CFt subunit ct; atpB = the gene for ATP synthetase CF~ subunit [3; atpE = the gene for ATP synthetase CF 1 subunit e; atpH = the gene for ATP synthetase CFo subunit III; psbA = gene for a 32,000 dalton photosystem II protein; rrs, rrl and rrf= 16S, 23S and 5S rRNA genes; trn ...

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Section: Probe Plasmid Hindlii Psti Pvullmentioning

confidence: 99%

Section: The Inverted Repeat and The Small Singlementioning

confidence: 99%

Section: Introducrlonmentioning

confidence: 99%