The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit β-globin DNA

Rb, Wallace; Johnson, Malcolm; Hirose, Tatsuro; Miyake, Toru; Kawashima, Eric; Itakura, Keiichi

doi:10.1093/nar/9.4.879

Cited by 470 publications

(184 citation statements)

References 20 publications

(14 reference statements)

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“…Transformation of E. eoti MC 1061 or HB101 was carried out according to Bolivar & Backman (1979) using RbCI. Colony hybridizations using synthetic oligodeoxynucleotides were performed as in Wallace et al (1981 a).…”

Section: Methodsmentioning

confidence: 99%

Hyperproduction of Polyhedrin-IGF II Fusion Protein in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus

Marumoto

Sato

Fujiwara

et al. 1987

Journal of General Virology

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SUMMARYA gene coding for insulin-like growth factor II (IGF II) was constructed from 16 oligodeoxynucleotides synthesized chemically and cloned into EcoRI-SalI sites of pBR322. In this gene an ATG codon for methionine was introduced for cleavage by CNBr at the beginning of mature IGF II. For expressing foreign genes, a new hostvector system, with Bombyx mori silkworm larvae as the host and B. mori nuclear polyhedrosis virus (BmNPV) as the vector, has been developed. BmNPV genomic DNA codes polyhedrin which is a major protein of inclusion bodies and is massproduced in infected silkworm larvae. We employed this polyhedrin production system to obtain a large yield of a foreign gene product. The coding region of the carboxyterminal half of polyhedrin was removed and the remainder was ligated with the IGF II gene in phase to create a fusion protein gene consisting of the coding region of the amino-terminal half of polyhedrin and the IGF II gene. This fusion protein gene was combined in a plasmid with the promoter and 5' and 3' flanking regions of the polyhedrin gene. The resulting plasmid and the wild-type BmNPV genomic DNA were cotransfected into BM-N cells, and a recombinant virus was isolated by the limiting dilution method. The silkworm larvae infected with the recombinant virus produced 3-6 mg of the fusion protein per larva and the infected BM-N cells produced 0-3 mg per ml of culture. IGF II was released from the fusion protein produced by BM-N cells infected with the recombinant virus by CNBr treatment, purified by extraction with guanidine-HC1, column chromatography and HPLC and the correct amino-terminal amino acid sequence confirmed. INTRODUCTIONIn order to express foreign genes, new host vector systems employing insect viruses and insect cell lines or larval bodies have recently been developed. Recombinant vaccinia virus was created to produce Sindbis virus proteins in Aedes albopictus mosquito cell lines (Franke & Hruby, 1985).

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Section: Methodsmentioning

confidence: 99%

Hyperproduction of Polyhedrin-IGF II Fusion Protein in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus

Marumoto

Sato

Fujiwara

et al. 1987

Journal of General Virology

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“…The foundation for clinical use of this strategy was the finding in 1981 (Wallace et al, 1981) that hybridization conditions could be found that allowed a 14-base-pair oligonucleotide to bind when perfectly matched and not bind when a single base pair mismatch occurred in the duplex. This system has been used and developed extensively up to the present.…”

Section: Mutation Detection Methods Where the Mutation Position Is Knmentioning

confidence: 99%

Detection of single base changes in nucleic acids

Cotton

1989

Biochemical Journal

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“…2), following the suppliers' recommendations (Biihringer Mannheim ; BRL; Pharmacia). The resulting fragments were separated on a 1 % (w/v) agarose gel, blotted onto nylon filters (Pall) and hybridized to the [~~~PIATP-end-labelled oligonucleotide probes (Wallace et al, 1979(Wallace et al, , 1981Conner et al, 1983).…”

Section: U B Gobel a G E I S E R A N D E J S T A N B R I Dmentioning

confidence: 99%

Oligonucleotide Probes Complementary to Variable Regions of Ribosomal RNA Discriminate between Mycoplasma Species

Göbel

Geiser²,

Stanbridge³

1987

Microbiology

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On the basis of information from computer-assisted sequence comparison of the Mycoplasma pneumoniae 16s ribosomal RNA (rRNA) sequences with sequences from various other mycoplasmal and bacterial species, we constructed M. pneumoniae-specific oligonucleotide probes complementary to variable regions in the 16s rRNA molecule. Using a DNA/RNA dot blot hybridization procedure, it was possible to detect less than 1 x lo3 mycoplasmas. This test is a most sensitive assay for species-specific detection of bacteria. It can easily be adapted for detection and identification of other bacterial species and may have wide medical and industrial application.

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The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit β-globin DNA

Cited by 470 publications

References 20 publications

Hyperproduction of Polyhedrin-IGF II Fusion Protein in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus

Hyperproduction of Polyhedrin-IGF II Fusion Protein in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus

Detection of single base changes in nucleic acids

Oligonucleotide Probes Complementary to Variable Regions of Ribosomal RNA Discriminate between Mycoplasma Species

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