Cloning of an almost full-length chicken conalbumin double-stranded cDNA

M. Cochet; F. Perrin; F. Gannon; A. Krust; P. Chambon; G. S. Mcknight; D. C. Lee; K. E. Mayo; R. Palmiter

doi:10.1093/nar/6.7.2435

Cloning of an almost full-length chicken conalbumin double-stranded cDNA

M. Cochet^*, F. Perrin, F. Gannon, A. Krust, P. Chambon, G. S. Mcknight, D. C. Lee, K. E. Mayo, R. Palmiter

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

44 Scopus citations

Abstract

Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by bluntend ligation into the SalI site of plasmid pBR322 which had been repaired with DNA polymerase I to create TaqI sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.

Original language	English (US)
Pages (from-to)	2435-2452
Number of pages	18
Journal	Nucleic acids research
Volume	6
Issue number	7
DOIs	https://doi.org/10.1093/nar/6.7.2435
State	Published - Jun 11 1979

Funding

We thank Drs. J.P. LePennec and P. Gerlinger for gifts of EcoRI, BamHI and HindHI restriction enzymes, and Dr. M. Bel-lard for a gift of nuclease SI. We thank the Viral Cancer Program, National Cancer Institute (Or. Beard) for gifts of avian myeloblastosis virus reverse transcriptase and Dr. A. Kornberg and D. Brutlag for gifts of purified E.coli DNA polymerase I. The excellent technical assistance of Mrs. C. Wasylyk, E. Sitt-ler and of Mr. J.M. G a m i e r , B. Boulay and E. Taubert is greatly acknowledged. Electron microscopy facilities were made available to F. Perrin in Strasbourg by the CNRS and the INSERM. This work was supported in Strasbourg by grants to P. Chambon from the INSERM (CRT 76.5.462 and 76.5.468), the CNRS (ATP 2117) and the Fondation pour la Recherche Medicale Frangaise, and in Seattle by grants to R. Palmiter from the National Institutes of Health (GM-15731 and HD 09172). Dr. C. Lee is supported by the Research Service Award GM-07270 from the National Institutes of Health. G.S. McKnight and R.D. Palmiter are Associate Investigator and Investigator of the Howard Hughes Medical Institute, respectively-

ASJC Scopus subject areas

Genetics

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10.1093/nar/6.7.2435

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title = "Cloning of an almost full-length chicken conalbumin double-stranded cDNA",

abstract = "Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by bluntend ligation into the SalI site of plasmid pBR322 which had been repaired with DNA polymerase I to create TaqI sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.",

author = "M. Cochet and F. Perrin and F. Gannon and A. Krust and P. Chambon and Mcknight, {G. S.} and Lee, {D. C.} and Mayo, {K. E.} and R. Palmiter",

note = "Funding Information: We thank Drs. J.P. LePennec and P. Gerlinger for gifts of EcoRI, BamHI and HindHI restriction enzymes, and Dr. M. Bel-lard for a gift of nuclease SI. We thank the Viral Cancer Program, National Cancer Institute (Or. Beard) for gifts of avian myeloblastosis virus reverse transcriptase and Dr. A. Kornberg and D. Brutlag for gifts of purified E.coli DNA polymerase I. The excellent technical assistance of Mrs. C. Wasylyk, E. Sitt-ler and of Mr. J.M. G a m i e r , B. Boulay and E. Taubert is greatly acknowledged. Electron microscopy facilities were made available to F. Perrin in Strasbourg by the CNRS and the INSERM. This work was supported in Strasbourg by grants to P. Chambon from the INSERM (CRT 76.5.462 and 76.5.468), the CNRS (ATP 2117) and the Fondation pour la Recherche Medicale Frangaise, and in Seattle by grants to R. Palmiter from the National Institutes of Health (GM-15731 and HD 09172). Dr. C. Lee is supported by the Research Service Award GM-07270 from the National Institutes of Health. G.S. McKnight and R.D. Palmiter are Associate Investigator and Investigator of the Howard Hughes Medical Institute, respectively-",

year = "1979",

month = jun,

day = "11",

doi = "10.1093/nar/6.7.2435",

language = "English (US)",

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AU - Cochet, M.

AU - Perrin, F.

AU - Gannon, F.

AU - Krust, A.

AU - Chambon, P.

AU - Mcknight, G. S.

AU - Lee, D. C.

AU - Mayo, K. E.

AU - Palmiter, R.

N1 - Funding Information: We thank Drs. J.P. LePennec and P. Gerlinger for gifts of EcoRI, BamHI and HindHI restriction enzymes, and Dr. M. Bel-lard for a gift of nuclease SI. We thank the Viral Cancer Program, National Cancer Institute (Or. Beard) for gifts of avian myeloblastosis virus reverse transcriptase and Dr. A. Kornberg and D. Brutlag for gifts of purified E.coli DNA polymerase I. The excellent technical assistance of Mrs. C. Wasylyk, E. Sitt-ler and of Mr. J.M. G a m i e r , B. Boulay and E. Taubert is greatly acknowledged. Electron microscopy facilities were made available to F. Perrin in Strasbourg by the CNRS and the INSERM. This work was supported in Strasbourg by grants to P. Chambon from the INSERM (CRT 76.5.462 and 76.5.468), the CNRS (ATP 2117) and the Fondation pour la Recherche Medicale Frangaise, and in Seattle by grants to R. Palmiter from the National Institutes of Health (GM-15731 and HD 09172). Dr. C. Lee is supported by the Research Service Award GM-07270 from the National Institutes of Health. G.S. McKnight and R.D. Palmiter are Associate Investigator and Investigator of the Howard Hughes Medical Institute, respectively-

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N2 - Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by bluntend ligation into the SalI site of plasmid pBR322 which had been repaired with DNA polymerase I to create TaqI sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.

AB - Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by bluntend ligation into the SalI site of plasmid pBR322 which had been repaired with DNA polymerase I to create TaqI sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.

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JF - Nucleic acids research

IS - 7

ER -

Cloning of an almost full-length chicken conalbumin double-stranded cDNA

Abstract

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