Studies on transfer ribonucleic acids and related compounds. XXI. Synthesis and properties of guanine rich fragments from E. coli tRNAfMet 5'-end.

Ohtsuka, Eiko; Nakagawa, Eiko; Tanaka, Toshiki; Markham, A.F.; Ikehara, Morio

doi:10.1248/cpb.26.2998

Cited by 16 publications

(5 citation statements)

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“…3 ml) was added. The mixture was stirred for N2-Isobutyryl-5'-O-(4-methoxytrityl)-2'-O-methylguanosine (13) [3d] N2-Isobutyryl-5'-O-(4-methoxytrityl)guanosine (12, 6.0 g, 9.6 mmol) [45] and SnCl2 (0.38 g, 2 mmol) was dissolved in DMF (480 ml) and cooled in an ice-bath. To the stirred solution, diazomethane (1,2-dimethoxyethane solution, 25 ml) [16] was added in portions over a period of 3 hr.…”

Section: Tablementioning

confidence: 99%

Synthesis and hybridization studies on two complementary nona(2'-O-methyl)ribonucleotides

et al. 1987

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2'-O-Methyl derivatives of the common ribonucleosides except for guanosine were synthesized via the 2'-O-methylation of appropriately-protected nucleosides with CH3I in the presence of Ag2O. The 2'-O-methylguanosine derivative was prepared by the monomethylation of a 2',3'-cis-diol system with diazomethane. These derivatives were converted to protected 2'-O-methylribonucleoside 3'-phosphates and used for oligonucleotide synthesis on polymer supports. Thus, oligo(2'-O-methyl-ribonucleotides) having the sequence identical to the consensus sequence of the 5'-splice junction CAGGUAAGU and its complement were synthesized in a stepwise manner using the phosphotriester method. Thermal stabilities (Tm's) of the duplex of these 2'-O-methyl ribo-oligomers and eight related duplexes containing ribo- or deoxyribo-oligomers were examined. It was found that the 2'-O-methyl oligoribonucleotides can be utilized as an alternative to an oligoribonucleotide probe in RNA hybridizations as the hybrid formed has a high, or a higher Tm, the probe is much easier to synthesize and it is less likely to be enzymatically degraded.

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Section: Tablementioning

confidence: 99%

Synthesis and hybridization studies on two complementary nona(2'-O-methyl)ribonucleotides

et al. 1987

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“…We began by synthesizing terminal fragments of the tRNA (11)(12)(13)(14)(15)(16) and examined the ability of RNA ligase (17)(18)(19) to join these fragments. The 5'-terminal icosanucleotide (20), the tetradecanucleotide (bases 21-34) (21), and the 3'-heptadecanucleotide (22) have been obtained by this method.…”

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confidence: 99%

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

Ohtsuka¹,

Tanaka²,

Tanaka³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNA; except that it lacks the base modifications present in the E. coli tRNA. This was achieved by enzymatic joining of chemically synthesized oligonucleotides with chain lengths.of3-10 which were synthesized by the phosphodiester or phosphotriester method. First, quarter molecules of tRNA were constructed by joining of chemically synthesized fragments with RNA ligase. The 5'-quarter molecule (bases 1-20) served as an acceptor in joining reactions with the 3',5'-bisphosphorylated donor molecule (bases 21-34). The 5'-half molecule thus obtained was treated with phosphatase andjoined to the 3'-half molecule which was prepared by ligation of the other quarter molecules (bases 35-60, acceptor; bases 61-77, donor) followed by 5'-phosphorylation with polynucleotide kinase. The synthetic tRNA was characterized by oligonucleotide pattern and was partially active in aminoacylation with E. coli methionyl-tRNA synthetase.Chemical synthesis ofnucleic acids has been a challenging problem in organic chemistry since the structure ofthe nucleic acids was elucidated. Chemical methods to synthesize short ribo-and deoxyribopolynucleotides with defined sequences were established in early 1960s, and those oligonucleotides were important in the elucidation of the genetic code (1). Discovery of DNA ligase allowed the synthesis of bihelical DNAs from chemically synthesized deoxyribopolynucleotides. With this chemical-enzymatic method the genes for yeast alanine tRNA (2) and Escherichia coli tyrosine tRNA precursor (3) have been synthesized; the latter was the first synthetic functional DNA molecule. Genes for peptides have also been synthesized by the same approach, and the methods for joining double-stranded DNA pieces with protruding ends have been used in various recently developed reactions for genetic manipulations.Although tRNAs are the smallest nucleic acids with unique functions, their synthesis has been difficult until recently, mainly because of the lack ofgood synthetic methods for larger .oligoribonucleotides as well as a lack ofjoining enzymes. After the primary structure of yeast alanine tRNA had been determined (4), the nona-and hexanucleotide corresponding to the terminal sequence of this tRNA were synthesized by phosphodiester block condensation. These fragments in turn were used to form reconstituted molecules with natural tRNA fragments derived by RNase digestions. However, aminoacylation was not possible because the synthetic fragments were too small to form sufficiently stable complexes for recognition by the alanyl-tRNA synthetase (5). The discovery of RNA ligase (6) and its ability to join single-stranded oligoribonucleotides (7) made it possible to elongate synthetic RNA fragments to yield larger molecules such as tRNAs.The initiator methionine tRNA of prokaryotes has a special role in protein biosynthesis, which. manifests itself in several unique properties of that tRNA (8). It was also the subject of detailed modif...

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“…In Basic Protocol 4, guanosine is alkylated after protection of its exocyclic amine with an isobutyryl group (Ohtsuka et al, 1978). Subsequent conversion to its dibutylstannylene derivative and alkylation in the absence of a catalyst leads to exclusive production of the 2′-O-(2-nitrobenzyloxymethyl) isomer.…”

Section: -Nitrobenzyloxymethyl Protectionmentioning

confidence: 99%

2′‐Hydroxyl‐Protecting Groups that are Either Photochemically Labile or Sensitive to Fluoride Ions

Miller

Schwartz

Gough

2000

CP Nucleic Acid Chemistry

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Protected ribonucleotide monomers are more difficult to obtain than their 2'-deoxy counterparts because of the need to protect the 2'-hydroxy function. This unit describes the stepwise preparation of suitably 2'-protected ribonucleosides using two protecting groups: 2-nitrobenzyloxymethyl (NBOM) and tert-butyldimethylsilyl (TBDMS). In addition, details are given for protecting the 5'-hydroxyl and the nucleobase, yielding nucleosides that are easily converted to phosphoramidite or H-phosphonate derivatives for automated oligoribonucleotide synthesis.

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Studies on transfer ribonucleic acids and related compounds. XXI. Synthesis and properties of guanine rich fragments from E. coli tRNAfMet 5'-end.

Cited by 16 publications

References 0 publications

Synthesis and hybridization studies on two complementary nona(2'-O-methyl)ribonucleotides

Synthesis and hybridization studies on two complementary nona(2'-O-methyl)ribonucleotides

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

2′‐Hydroxyl‐Protecting Groups that are Either Photochemically Labile or Sensitive to Fluoride Ions

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