Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA

R. Brousseau; R. Scarpulla; W. Sung; H. M. Hsiung; S. A. Narang; Ray Wu

doi:10.1016/0378-1119(82)90144-5

Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA

R. Brousseau, R. Scarpulla, W. Sung, H. M. Hsiung, S. A. Narang, Ray Wu

Cell & Developmental Biology

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

35 Scopus citations

Abstract

To form a 258-bp sequence coding for human proinsulin, 41 synthetic deoxyribo-oligonucleotide fragments of 11 to 15 nucleotides in length were assembled by enzymatic methods. The coding sequence is preceded by ATG and followed by TGA for translation start and stop signals, and terminated in an EcoRI and a BamHI recognition sequence. The complete synthetic sequence was ligated to a plasmid and cloned in Escherichia coli. The cloned DNA was shown to have the correct human proinsulin coding sequence.

Original language	English (US)
Pages (from-to)	279-289
Number of pages	11
Journal	Gene
Volume	17
Issue number	3
DOIs	https://doi.org/10.1016/0378-1119(82)90144-5
State	Published - Mar 1982

Funding

We wish to thank Douglas H. Smith and J. Michniewicz for excellent technicaals sistance and Li-He Guo for valuable help with the sequence analysis of the cloned proinsulin gene. The work was supported by research grant AM21801 from the National Institute of Health, U.S. Public Health Service, from the National Research Council of Canada (NRCC NO 19599)a nd a fellowship grant DRG-323F (to R.C.S.) from the Damon Runyon-Walter Winchell Cancer Fund.

Keywords

Recombinant DNA
plasmid vectors
start and stop signals

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(82)90144-5

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title = "Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA",

abstract = "To form a 258-bp sequence coding for human proinsulin, 41 synthetic deoxyribo-oligonucleotide fragments of 11 to 15 nucleotides in length were assembled by enzymatic methods. The coding sequence is preceded by ATG and followed by TGA for translation start and stop signals, and terminated in an EcoRI and a BamHI recognition sequence. The complete synthetic sequence was ligated to a plasmid and cloned in Escherichia coli. The cloned DNA was shown to have the correct human proinsulin coding sequence.",

keywords = "Recombinant DNA, plasmid vectors, start and stop signals",

author = "R. Brousseau and R. Scarpulla and W. Sung and Hsiung, {H. M.} and Narang, {S. A.} and Ray Wu",

note = "Funding Information: We wish to thank Douglas H. Smith and J. Michniewicz for excellent technicaals sistance and Li-He Guo for valuable help with the sequence analysis of the cloned proinsulin gene. The work was supported by research grant AM21801 from the National Institute of Health, U.S. Public Health Service, from the National Research Council of Canada (NRCC NO 19599)a nd a fellowship grant DRG-323F (to R.C.S.) from the Damon Runyon-Walter Winchell Cancer Fund.",

year = "1982",

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doi = "10.1016/0378-1119(82)90144-5",

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T1 - Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA

AU - Brousseau, R.

AU - Scarpulla, R.

AU - Sung, W.

AU - Hsiung, H. M.

AU - Narang, S. A.

AU - Wu, Ray

N1 - Funding Information: We wish to thank Douglas H. Smith and J. Michniewicz for excellent technicaals sistance and Li-He Guo for valuable help with the sequence analysis of the cloned proinsulin gene. The work was supported by research grant AM21801 from the National Institute of Health, U.S. Public Health Service, from the National Research Council of Canada (NRCC NO 19599)a nd a fellowship grant DRG-323F (to R.C.S.) from the Damon Runyon-Walter Winchell Cancer Fund.

PY - 1982/3

Y1 - 1982/3

N2 - To form a 258-bp sequence coding for human proinsulin, 41 synthetic deoxyribo-oligonucleotide fragments of 11 to 15 nucleotides in length were assembled by enzymatic methods. The coding sequence is preceded by ATG and followed by TGA for translation start and stop signals, and terminated in an EcoRI and a BamHI recognition sequence. The complete synthetic sequence was ligated to a plasmid and cloned in Escherichia coli. The cloned DNA was shown to have the correct human proinsulin coding sequence.

AB - To form a 258-bp sequence coding for human proinsulin, 41 synthetic deoxyribo-oligonucleotide fragments of 11 to 15 nucleotides in length were assembled by enzymatic methods. The coding sequence is preceded by ATG and followed by TGA for translation start and stop signals, and terminated in an EcoRI and a BamHI recognition sequence. The complete synthetic sequence was ligated to a plasmid and cloned in Escherichia coli. The cloned DNA was shown to have the correct human proinsulin coding sequence.

KW - Recombinant DNA

KW - plasmid vectors

KW - start and stop signals

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JO - Gene

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Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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