Simplifications in the synthesis of short oligonucleotide blocks

Broka, Chris A.; Hozumi, Toyoharu; Arentzen, René; Itakura, Keiichi

doi:10.1093/nar/8.22.5461

Cited by 73 publications

(25 citation statements)

References 8 publications

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“…,B2mIII was synthesized by a solidphase method (17). All oligonucleotides were purified by highperformance liquid chromatography on an ion-exchange (Du Pont Permaphase AAX) column (5 (21).…”

Section: Methodsmentioning

confidence: 99%

Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin.

Suggs¹,

Rb²,

Hirose³

et al. 1981

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

372

152

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We have synthesized two sets of 15-base-long oligodeoxyribonucleotides corresponding to all possible coding sequences for a small portion of human fl2-microglobulin. Labeled oligonucleotides were used as hybridization probes to screen bacterial clones containing cDNA sequences primed with oligo(dT) and inserted into the plasmid vector pBR322. One A-microglobulin cDNA clone was detected in the 535 bacterial plasmid clones that were screened. The clone has been characterized by blotting and nucleotide sequence analysis. The cloned P2-microglobulin sequence contains 217 base pairs of the 3' untranslated region of the mRNA and 328 base pairs (97%) of the coding region.

show abstract

Section: Methodsmentioning

confidence: 99%

Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin.

Suggs¹,

Rb²,

Hirose³

et al. 1981

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

372

152

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“…Several laboratories have reported the purification of peptides possessing the above activities (3)(4)(5) made by the solid-phase triester method (11). The following sequences were made and will be designated as ANF primer…”

mentioning

confidence: 99%

Molecular cloning and characterization of DNA sequences encoding rat and human atrial natriuretic factors.

Zivin¹,

Condra²,

Dixon³

et al. 1984

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

109

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A cDNA copy of the message encoding rat atrial natriuretic factor (ANF) has been cloned in Escherichia coli, and its nucleotide sequence was determined. ANF appears to be synthesized as a larger precursor, atrial pronatriodilatin.The cDNA has an open reading frame potentially encoding a protein of 152 amino acids, of which the first 24 amino acids strongly resemble a signal sequence. This is followed by a sequence with 80% homology to a second vasoactive protein, porcine cardiodilatin. The ANF peptide is contained in the COOH-terminal portion of the protein. The DNA sequence corresponding to human ANF is also presented and displays a high degree of homology to its rat counterpart. These data provide further evidence for the expression in cardiac atria of a multifactor system that may contribute to the regulation of blood pressure and extracellular fluid volume.It has been postulated for many years that cardiac atria possess sensors that are important in detecting changes in extracellular fluid volume (1). Such a system could respond and compensate for these changes through release of a humoral substance.The existence of such an atrially produced substance, which enhances urinary sodium excretion and, therefore, might be involved in regulation of extracellular fluid volume, was recently demonstrated by de Bold et al. (2). When a partially purified extract of rat cardiac atria was injected into anesthetized rats, a large increase of urine flow and urinary sodium excretion was observed. This relatively crude extract, which possessed the appropriate characteristics for an endogenous natriuretic substance, was named atrial natriuretic factor (ANF).In addition to its potent and natriuretic effects, it also was discovered that these extracts have potent smooth muscle relaxant activity. This property implies that they might have a potential role in regulating blood pressure as well as extracellular fluid volume.Several laboratories have reported the purification of peptides possessing the above activities (3-5). Seidah et al. (6) identified a sequence common to all of the active peptides, which when synthesized, show activity in animals similar to endogenous ANF.In this paper, we describe the isolation of a cDNA clone containing the sequence encoding this active ANF peptide. The clone spans nearly the entire rat ANF mRNA. Sequence analysis revealed that ANF was contained in a large precursor polypeptide, atrial pronatriodilatin (7), which also demonstrated homology to a second vasoactive peptide, cardiodilatin (8). Thus, the products of the ANF gene might act at several levels to control blood pressure and fluid volume.In addition, we have used the rat cDNA to isolate clones of the corresponding human gene and have found a high degree of sequence homology between rat and human ANF. MATERIALS AND METHODS

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“…Phosphodiesters (6) were prepared as described. 8 Comparison of time required for removal of aryl esters on an internucleotide phosphate…”

Section: Resultsmentioning

confidence: 99%

Studies on deoxyribonucleic acids and related compounds. V. Synthesis of pentadecanucleotide duplex containing the ideal Pribnow sequence of promoter by the phosphotriester method using a new phosphorylating reagent

et al. 1982

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A phosphorylating reagent o-chlorophenyl phosphoro-p-anisidochloridate was synthesized to phosphorylate the 3'-hydroxyl group of N, 5'-protected deoxynucleosides. These nucleotides served as 3'-terminal units for the synthesis of oligonucleotide blocks. By condensation of these oligonucleotide blocks the partially complementary deoxypentadecanucleotides dAGCTTATAATGC-TCG and dAGCTCGAGCATTATA, which contained the ideal Pribnow sequence TATAATG, were synthesized.

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Simplifications in the synthesis of short oligonucleotide blocks

Cited by 73 publications

References 8 publications

Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin.

Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin.

Molecular cloning and characterization of DNA sequences encoding rat and human atrial natriuretic factors.

Studies on deoxyribonucleic acids and related compounds. V. Synthesis of pentadecanucleotide duplex containing the ideal Pribnow sequence of promoter by the phosphotriester method using a new phosphorylating reagent

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