Ligase joining of oligodeoxythymidylate

Harvey, Clifford; Wright, Rosemary

doi:10.1021/bi00764a018

Cited by 16 publications

(6 citation statements)

References 11 publications

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“…25-50 bases in length, a result consistent with joining of single (dT)lo molecules onto members of the population of (rA)9.60 molecules. End-to-end joining of the (dT)lo to itself was not detected; it would have yielded discrete bands of n = 20, 30,... bases in length such as those shown in lane 9 of Figure 8 which were generated by T4 DNA ligase (12). Such a reaction for T4 RNA ligase would be unlikely under these conditions in any case based on the preceding section.…”

Section: Ligation Of 5'-32p-(dt)1n To Oligoriboadenylatesmentioning

confidence: 91%

Polymer-stimulated ligation: enhanced ligation of oligo- and polynucleotides by T4 RNA ligase in polymer solutions

Harrison

Zimmerman

1984

Nucl Acids Res

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The effects of macromolecular crowding were tested on several reactions catalyzed by T4 RNA ligase. The rate of cyclization of oligoriboadenylates was stimulated up to 10-fold by relatively high concentrations of several polymers (polyethylene glycol (PEG) 8000 or 20,000; bovine plasma albumin; Ficoll 70). In addition, higher concentrations of PEG 8000 or PEG 20,000 allowed the novel formation of large linear products from the oligoriboadenylates. Also stimulated by high concentrations of PEG 8000 were the rate at which T4 RNA ligase joined p(dT)10 to oligoriboadenylates and the rate at which the enzyme activated p(dT)n by transfer of an adenylyl moiety from ATP to the oligonucleotides. These results with T4 RNA ligase are compared to earlier studies on the effects of crowding on DNA ligases.

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Section: Ligation Of 5'-32p-(dt)1n To Oligoriboadenylatesmentioning

confidence: 91%

Polymer-stimulated ligation: enhanced ligation of oligo- and polynucleotides by T4 RNA ligase in polymer solutions

Harrison

Zimmerman

1984

Nucl Acids Res

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“…Micrococcal nuclease, venom phosphodiesterase and spleen phosphodiesterase were obtained from Worthington Biochemical Corporation. 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%

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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“…Enzymes. Polynucleotide kinase and polynucleotide ligase from bacteriophage T4 infected bacterial cells were the same as used previously (Harvey and Wright, 1972). Calf intestinal mucosa alkaline phosphatase (type VII) was obtained from Sigma Chemical.…”

Section: Methodsmentioning

confidence: 99%

“…Other Materials. The ATP used was obtained and purified as described earlier (Harvey and Wright, 1972). Crystalline bovine albumin was purchased from Schwarz-Mann.…”

Section: Methodsmentioning

confidence: 99%