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An optimized recipe for cloning of the polymerase chain reaction-amplified DNA inserts into plasmid vectors

100%

Topcu Z.

Acta Biochimica Polonica

2000

tom 47

nr 3

This study compares a number of parameters that are important in the ligation of the polymerase chain reaction-amplified DNA inserts into plasmid vectors and their efficient transformation to bacterial cells. The parameters covered were: T4 polynucleotide kinase treatment followed by either the large fragment of E. coli DNA polymerase or T4 DNA polymerase reactions, the amount of T4 DNA ligase, temperature and duration of ligation, molar ratio of insert to vector as well as the total DNA concentration. The results show that the T4 polynucleotide kinase-treated group without further enzymatic manipulation, at an insert to vector ratio of 3:1 gave the highest recombination efficiency when 10 µg/ml DNA and 20 units T4 DNA ligase were applied for ligation for 12 h at 4°C.

Controversial aspect of using GFP as a marker for the production of transgenic cattle

100%

Duszewska A. M., Lipinski D., Piliszek A., Slomski R., Plawski A., Wojdan J., Gawron W., Juzwa W., Zeyland J., Wenta-Muchalska E., Reklewski Z.

Polish Journal of Veterinary Sciences

2004

tom 07

nr 4

The objective of this study was to examine the feasibility of identification and selection of cattle embryos based on green fluorescence (GFP-positive) in order to obtain calves carrying an integrated transgene. The construct used (pbLGTNF-EGFP) contained the human tumor necrosis factor alpha (hTNFa) gene fused to the bovine betalactoglobulin promoter (bLG) in plasm id vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulac/blastocysts stage of which only five were GFP positive (one of them 100%, one-50%, three- 25%). All of the GFP positive embryos were transferred to recipients. Two calves were born: one after transfer of the 100% GFP positive embryo and the other after transfer of one of the 25% GFP positive embryos. Both animals were healthy with normal weight when compared to two control calves. The integration of pbLGTNF-EGFP in the host genome could not be detected in either of the calves, suggesting that GFP is an unreliable marker for preimplantation screening of embryos.

Soluble TNF-alpha receptor 1 encoded on plasmid vector and its application in experimental gene therapy of radiation-induced lung fibrosis

80%

Przybyszewska M., Miloszewska J., Rzonca S., Trembacz H., Pysniak K., Kotlarz A., Swoboda P., Zalewska M., Malecki M.

Archivum Immunologiae et Therapiae Experimentalis

2011

tom 59

nr 4

Ograniczanie wyników

1 Acta Biochimica Polonica

1 Archivum Immunologiae et Therapiae Experimentalis

1 Polish Journal of Veterinary Sciences

1 Duszewska A. M.

1 Gawron W.

1 Juzwa W.

1 Kotlarz A.

1 Lipinski D.

1 Malecki M.

1 Miloszewska J.

1 Piliszek A.

1 Plawski A.

1 Przybyszewska M.

1 Pysniak K.

1 Reklewski Z.

1 Rzonca S.

1 Slomski R.

1 Swoboda P.

1 Topcu Z.

1 Trembacz H.

1 Wenta-Muchalska E.

1 Wojdan J.

1 Zalewska M.

1 Zeyland J.

1 2011

1 2004

1 2000

Wyniki wyszukiwania

An optimized recipe for cloning of the polymerase chain reaction-amplified DNA inserts into plasmid vectors

Controversial aspect of using GFP as a marker for the production of transgenic cattle

Soluble TNF-alpha receptor 1 encoded on plasmid vector and its application in experimental gene therapy of radiation-induced lung fibrosis