Synthesis of deoxyoligonucleotides on a polymer support

Matteucci, Mark D.; Caruthers, M. H.

doi:10.1021/ja00401a041

Cited by 935 publications

(462 citation statements)

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“…This latter reaction is similar to that reported for arabinosyl adenosine (40) and various pyrimidine derivatives (41). Deprotection of the carbohydrate residue and subsequent preparation of the dimethoxytrityl phosphoramidite derivative 6 (Scheme 1), produced a compound suitable for incorporation 5637 I H2NNH2 into oligodeoxynucleotides using phosphite triester synthesis procedures (32). We have found that silver oxide oxidation of the hydrazino derivative i (Scheme 1) is a simpler and more convenient route to the 2-aminopurine chromophore than previously described hydrogenation (27) or elimination reactions (28,29).…”

Section: Resuitssupporting

confidence: 74%

“…We wish to report a new approach to the synthesis of the 2-aminopurine heterocycle and the synthesis of the corresponding phosphoramidite derivative which allows incorporation of the modified nucleobase into oligodeoxynucleotides using chemical phosphite triester synthesis procedures (31,32). We have prepared four 2-aminopurine containing decadeoxynucleotides and one dodecadeoxynucleotide containing such that the 2-aminopurine residue has either a complementary thymine or cytosine nucleobase.…”

Section: Introductionmentioning

confidence: 99%

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A new approach to the synthesis of a protected 2-aminopurine derivative and its incorporation into oligodeoxynucleotides containing the Eco RI and Bam HI recognition sites

et al. 1988

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A protected 2-aminopurine nucleoside suitable for incorporation into oligodeoxynucleotides using phosphite triester chemical synthesis procedures has been prepared via oxidation of a purine hydrazino derivative with silver (I) oxide. Five oligodeoxynucleotides containing Eco RI and Bam HI recognition sites have been prepared such that, in the double stranded form, the 2-aminopurine base has either a complementary thymine or cytosine nucleobase. The helix character and thermodynamic parameters for helix formation have been examined.

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Section: Resuitssupporting

confidence: 74%

Section: Introductionmentioning

confidence: 99%

A new approach to the synthesis of a protected 2-aminopurine derivative and its incorporation into oligodeoxynucleotides containing the Eco RI and Bam HI recognition sites

et al. 1988

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“…A synthetic oligonucleotide (5'-CGAGCTTG-GATCAGGTCTACGGTGA-3') from the structural Synechococcus PCC7942psbO gene was used as a probe in hybridization experiments. The oligonucleotide was synthesized with an Applied Biosystems DNA Synthesizer Model 380B using the phosphoramitide method [16,17]. OPC-purifcation of the oligonucleotide was carried out as recommended by Applied Biosystems.…”

Section: Dna Jilter Hybridizationsmentioning

confidence: 99%

Insertional inactivation of the psbO gene encoding the manganese stabilizing protein of photosystem II in the cyanobacterium Synechococcus PCC7942

1991

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A S~nechococcus PCC7942 mutant in which the psh0 gene was inactivated by insertion of a chloramphenicol interposon and which did not contain any detectable manganese stabilizing protein in immunoblot experiments, was constructed. Such a Synechococcus mutant was able to grow under photoautotrophic conditions. Isolated thylakoid membranes from the mutant required addition of CaClz and MnClz for photosynthetic O2 evolution, and the detectable L-amino acid oxidase activity in the isolated thylakoid membranes from the mutant was approximately four times higher than in wild-type thylakoids. The results are discussed with respect to our model suggesting that the water-oxidizing enzyme may have evolved from a flavoprotein with L-amino acid dehydrogenase/oxidase activity.

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“…DNA was sequenced by the dideoxy method, using Sequenase modified T7 DNA polymerase (30). Deoxyoligonucleotides were synthesized on an Applied Biosystems DNA synthesizer (model 380B), using the phosphite triester method (9,21 follows. Plasmid pALA621 was derived from the hydroxylamine-induced mutation rep-30, which changes C to T at P1 base 457 (8).…”

Section: Methodsmentioning

confidence: 99%

Critical sequences in the core of the P1 plasmid replication origin

Brendler¹,

Abeles²,

Austin³

1991

J Bacteriol

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The core of the P1 plasmid replication origin consists of a series of 7-bp repeats and a G+C-rich stretch. Methylation of the GATC sequences in the repeats is essential. Forty different single-base mutations in the region were isolated and assayed for origin function. A single-base change within any 7-bp repeat could block the origin, irrespective of whether GATC bases were affected. The repeats themselves were critical, but the short intervals between them were not. Mutations in the G+C-rich region showed it to be a spacer whose exact length is important but whose sequence can vary considerably. It maintains a precise distance between the 7-bp repeats and binding sites for the P1 RepA initiator protein. It may also serve as a clamp to limit strand separation during initiation.The prophage of bacteriophage P1 is an autonomous plasmid that can be maintained in its Escherichia coli host at a copy number as low as one to two per cell (28). In this respect, it mimics the stringent behavior of the host chromosome. The P1 plasmid origin has some structural similarities to oriC, the origin of the host chromosome. The study of these features should be informative as to how replication control can achieve accurate maintenance at such a low copy number. The P1 replicon consists of three contiguous elements. These are the origin of replication, oriR; the gene for the essential replication protein, repA; and the incA locus, which controls plasmid copy number (2). The copy control region is not essential to replication, but when it is deleted, the copy number goes up to about 8 to 10 per cell instead of the 1 to 2 found with the wild-type plasmid (27). This incA locus consists of a series of 19-bp repeats of a sequence that is a specific binding determinant for the plasmid RepA protein (1).The origin, oriR, is a sequence of approximately 250 bp ( Fig. 1) sequences, we isolated a large number of mutants of the region and tested their phenotypes. MATERIALS AND METHODSBuffers, media, enzymes, and reagents. Conditions for cell growth and buffers used for electrophoresis and DNA preparation were previously described (3,5,20

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Synthesis of deoxyoligonucleotides on a polymer support

Cited by 935 publications

References 1 publication

A new approach to the synthesis of a protected 2-aminopurine derivative and its incorporation into oligodeoxynucleotides containing the Eco RI and Bam HI recognition sites

A new approach to the synthesis of a protected 2-aminopurine derivative and its incorporation into oligodeoxynucleotides containing the Eco RI and Bam HI recognition sites

Insertional inactivation of the psbO gene encoding the manganese stabilizing protein of photosystem II in the cyanobacterium Synechococcus PCC7942

Critical sequences in the core of the P1 plasmid replication origin

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