Nucleoside phosphorothioates. IX. Use of phosphate-blocking groups in ligase joining of oligodeoxyribonucleotides

Cl, Harvey; Wright, Rosemary; Af, Cook; Dt, Maichuk; Al, Nussbaum

doi:10.1021/bi00726a005

Cited by 11 publications

(6 citation statements)

References 19 publications

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“…Polynucleotide kinase was obtained as previously described (7). Calf intestinal alkaline phosphatase was obtained from Sigma Chemical Company and analyzed as previously described (8). …”

Section: Materiais and Methodsmentioning

confidence: 99%

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Enzymatic multiplication of a chemically synthesized DNA fragment

Olson¹,

Gabriel²,

Michalewsky³

et al. 1975

Nucl Acids Res

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A synthetic DNA fragment of 19 residues was enlarged by the enzymatic addition of deoxyadenylate residues to its 3'-end with calf thymus terminal deoxynucleotidyl transferase. The 3'-terminus of this elongated DNA strand was blocked with 2',3'-dideoxyadenylate to prevent hydrolysis by the 3'-exonuclease function of E. coli DNA polymerase I. This elongated and 3'-blocked fragment was annealed to an oligomeric primer and used as a template for the synthesis of a complementary copy of the synthetic 19-mer. The product of such a repair synthesis was separated by gel filtration and analyzed by nearest neighbor techniques.All template strands were copied with complgte repair in over 90% of the chains. Facile recovery of the elongated template by virtue of its size permitted repetition of the copy process, thus allowing accumulation of the desired strand.

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“…Polynucleotide kinase was obtained as previously described (7). Calf intestinal alkaline phosphatase was obtained from Sigma Chemical Company and analyzed as previously described (8). …”

Section: Materiais and Methodsmentioning

confidence: 99%

“…The nucleotide ratios found after a nearest neighbor analysis were close to those expected (Table 3B). (8).…”

Section: A-t-t-a-a) and D(pa-a-g-a-c-a-g-c-a-t-a-t)(l) The Preparatimentioning

confidence: 99%

Enzymatic multiplication of a chemically synthesized DNA fragment

Olson¹,

Gabriel²,

Michalewsky³

et al. 1975

Nucl Acids Res

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“…The 5'-termini of Fragments (Fig. 1) were 32 labeled with P in the polynucleotide kinase reaction after dephosphorylation with calf intestinal alkaline phosphatase (14). Isolation was carried out on Sephadex G-75 or G-100,…”

Section: Joining Of Oligomers By T 4polynucleotide Ligasementioning

confidence: 99%

“…both superfine, in jacketed columns at 700. The kinetics of joining were followed with the Norit assay (14). The joined Fragments were separated on the gel column with 0.1 M triethylammonium bicarbonate pH 7.6 (pH 8.3 after heating).…”

Section: Auggaaacugcggcugcuaaauuugaaagacagcauauggauuaamentioning

confidence: 99%

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Construction of a double-stranded deoxyribonucleotide sequence of 45 base pairs designed to code for S-peptide_2-14of bovine ribonuclease A

et al. 1975

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Synthese eines Strukturgens für das Peptidhormon Angiotensin II, Teil 9. Ligation der Teilsequenzen zum kompletten Doppelstrang

Blöcker

Frank

Köster

1978

Justus Liebigs Ann. Chem.

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Die Ligation der sieben Teilsequenzen+zum kompletten 33 Basenpaare umfassenden Doppelstrang einer Angiotensin-11-DNS mit Hilfe von T4 Polynucleotidligase wird beschrieben. Die Ligationsreaktion wird durch einen rnodifizierten Norittest verbunden mit einem Abbau von [y3*P]ATP mit Anorganischer Pyrophosphatase aus Backerhefe verfolgt. Diese neue Analysentechnik macht eine zeitraubende und verlustreiche Reinigung der mit T4 Polynucleotidkinase und [y3*P]ATP phosphorylierten Teilsequenzen uberfliissig und erlaubt es, die DNS-Synthese mehrstufig und ohne Zwischenreinigung durchzufuhren. Synthesis of a Structural Gene for the Peptide Hormone Angiotensin 11, Part 9l). -Ligation of the Partial Sequences to the Complete DuplexLigation of the seven partial sequences by T4 polynucleotide ligase to give the total duplex of an angiotensin I1 DNA with 33 base pairs is described. The ligation reaction was monitored by a modified Norit assay combined with the degradation of [Y-~'P]ATP by inorganic pyrophosphatase from baker's yeast. This new analytical technique makes it unnecessary to purify the segments after phosphorylation with T4 polynucleotide kinase and [y3'P]ATP by time-consuming and yielddecreasing methods. Moreover, this technique permits the synthesis of double helical DNA in a multiple step reaction without any intermediate purification. EinleitungNachdem in den voranstehenden Arbeiten (Teile 1 -8 dieser Reihe) die Synthese der sieben Teilsequenzen und deren Sequenzierung beschrieben wurden, wird in dieser Verdffentlichung uber ihre Kopplung zurn totalen Doppelstrang einer DNS fur das Peptidhorrnon Angiotensin I1 (Abbildung 1) berichtet. Eine Kurzmitteilung hieriiber ist bereits erschienen2). Anschlierjend wurde iiber die Synthese der Lactose-Operator-DNS3s4), einer DNS fur den Tyrosin-tRNS-Vorlaidfer aus E. c01i5), uber eine DNS fur Korrespondenz bitte an diesen Autor richten. H. Blocker und H.

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Nucleoside phosphorothioates. IX. Use of phosphate-blocking groups in ligase joining of oligodeoxyribonucleotides

Cited by 11 publications

References 19 publications

Enzymatic multiplication of a chemically synthesized DNA fragment

Enzymatic multiplication of a chemically synthesized DNA fragment

Construction of a double-stranded deoxyribonucleotide sequence of 45 base pairs designed to code for S-peptide_2-14of bovine ribonuclease A

Synthese eines Strukturgens für das Peptidhormon Angiotensin II, Teil 9. Ligation der Teilsequenzen zum kompletten Doppelstrang

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Nucleoside phosphorothioates. IX. Use of phosphate-blocking groups in ligase joining of oligodeoxyribonucleotides

Cited by 11 publications

References 19 publications

Enzymatic multiplication of a chemically synthesized DNA fragment

Enzymatic multiplication of a chemically synthesized DNA fragment

Construction of a double-stranded deoxyribonucleotide sequence of 45 base pairs designed to code for S-peptide2-14of bovine ribonuclease A

Synthese eines Strukturgens für das Peptidhormon Angiotensin II, Teil 9. Ligation der Teilsequenzen zum kompletten Doppelstrang

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Construction of a double-stranded deoxyribonucleotide sequence of 45 base pairs designed to code for S-peptide_2-14of bovine ribonuclease A