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A chemical-enzymatic synthesis of 271- and 286-bp DNA duplexes, each of which contains the entire sequence coding for human proinsulin has been accomplished. In addition to the coding sequence, the 271-bp fragment carries translation initiation and termination signals plus EcoRI-HindIII restriction enzyme sites for insertion into an appropriate plasmid vector. The 286-bp fragment also contains a Shine-Dalgarno (SD) sequence preceding an ATG codon. Employing the 286-bp polynucleotide, the 568-bp tandem proinsulin gene has been obtained. The synthesis of these DNA fragments involved preparation of 42 oligonucleotides by a rapid N-methylimidazolide phosphotriester method and enzymatic conversion of the oligonucleotides into the gene subfragments, which were cloned separately and fused to yield the desired DNAs coding for proinsulin. The proinsulin gene fragments were cloned in Escherichia coli and shown to have the correct sequences.
Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
November 2010,
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Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
March 1982,
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Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
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Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
September 1986,
Zhonghua yi xue za zhi,
Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
May 1986,
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Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
May 1986,
Chemical & pharmaceutical bulletin,
Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
August 1980,
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Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
October 2003,
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica,
Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
June 1979,
Hoppe-Seyler's Zeitschrift fur physiologische Chemie,
Y A Ovchinnikov, and
V A Efimov, and
I N Ivanova, and
S V Reverdatto, and
N P Skiba, and
O G Chakhmakhcheva
January 1973,
Chemische Berichte,
