Cloning of the DNA fragments reflecting the open reading frame I and II of the I-18 C gene of Chironomus tentans. II. Preparation of the intermediate vector - bluescript plasmid for ligation with the inserts

• Autorzy: Borowicz B P
• Języki publikacji: EN

Abstrakty

EN

The technology used for the cloning of the open reading frame (ORF) I and II reflecting DNA fragments of the I-18 C gene in regard to the preparation of the intermediate vector i.e. the bluescript for the ligation with the ORF I and II reflecting DNA fragments, is presented. The main steps include the Eco RV digestion of the bluescript plasmid, the phenol / methylene chloride extraction of the plasmid, dephosphorylation of the bluescript plasmid and finally its extraction with the phenol and ethanol precipitation.

PL

Przedstawiono technologię, którą zastosowano do klonowania fragmentów DNA odzwierciedlających translacyjną otwartą ramę odczytu I i II genu I-18 C Chironomus tentans w zakresie: przygotowania wektora pośredniego - plazmidu blueskrypt do ligacji z wymienionymi sekwencjami wstawek. Główne etapy obejmują: trawienie plazmidu blueskrypt enzymem Eco RV, ekstrakcję plazmidu fenolem/ chlorkiem metylenu, defosforylację plazmidu blueskrypt, ostatecznie jego oczyszczenie poprzez ekstrakcję fenolową i precypitację etanolem.

Słowa kluczowe

EN

cloning; I-18 C gene; plasmid; Chironomus tentans; polymerase chain reaction; DNA fragment

• Wydawca: -
• Czasopismo: Journal of Plant Protection Research
• Rocznik: 1999
• Tom: 39
• Numer: 1
• Opis fizyczny: p.46-49

Twórcy

autor

Borowicz B P

• Institute of Plant Protection, Miczurina 20, 60-318 Poznan, Poland

Bibliografia

• 1. Borowicz B.P. 1995. Isolation of the DNA fragment reflecting the Open Reading Frame I of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. I. The technology for the preparation of the samples (in English). Prace Nauk. Inst. Ochr. Roślin 36 (1/2): 69-76.
• 2. Borowicz B.P. 1995. Isolation of the DNA fragment reflecting the Open Reading Frame I of the I-18 C gene of Chironomus tentans by the Polymerase Chain Rcaction. II. The technology required to obtain the DNA fragment (in English). Prace Nauk. Inst. Ochr. Roślin 36 (1/2): 77-82.
• 3. Borowicz B.P. 1995. Isolation of the DNA fragment reflecting the Open Reading Frame I of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. III. The effect of the applied technologies (in English). Prace Nauk. Inst. Ochr. Roślin 36 (1/2): 83-93.
• 4. Borowicz B.P. 1996. Isolation of the DNA fragment reflecting the Open Reading Frame II of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. I. The technology for the preparation of the primers. J. Plant Protection Res., 37 (1/2): 138-144.
• 5. Borowicz B.P. 1996. Isolation of the DNA fragment reflecting the Open Reading Frame II of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. II. The technology for the preparation of the template. J. Plant Protection Res., 37, (1/2): 145-148.
• 6. Borowicz B.P. 1996. Isolation of the DNA fragment reflecting the Open Reading Frame II of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. III. The DNA amplification technology. J. Plant Protection Res., 37 (1/2): 149-155.
• 7. Borowicz B.P. 1996. Isolation of the DNA fragment reflecting the Open Reading Frame II of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. IV. The product of the applied technologies. J. Plant Protection Res., 37 (1/2): 156-164.
• 8. Borowicz B.P. 1996. Isolation of the DNA fragment reflecting the Open Reading Frame II of the I-18 C gene of Chironomus tentans by the Polymerase Chain Reaction. V. Different mechanisms of regulation regarding the Open Reading Frames. J. Plant Protection Res., 37 (1/2): 165-171.
• 9. Lezzi M., Lutz B., Dorsch-Haesler K. 1989 b. The ecdysterone-controlled chromosome region I-18 C in C. tentans: from cytology to molecular biology. Acta Biol. Debr. Oecol. Hung., 2: 129-139.
• 10. Sambrook J., Fritsch E.F., Maniatis T. 1989. Molecular Cloning. A laboratory manual. Second Edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, Books: 1-3.
• 11. Sambrook J., Fritsch E.F., Maniatis T. 1989. Plasmid Vectors (chapter 1) in Molecular Cloning. A laboratory manual. Second Edition. Cold Spring Habror Laboratory Press, Cold Spring Harbor, New York, Book 1:1.2-1.110.
• 12. Sambrook J., Fritsch E.F., Maniatis T. 1989. Expression of Cloned Genes in Escherichia coli (chapter 17) in Molecular Cloning. A laboratory manual. Second Edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, Book 3: 17.2-17.44.
• 13. Studier F.W., Rosenberg A.H., Dunn J.J., Dubendorff J.W. 1990. Use of T7 RNA Polymerase to direct expression of cloned genes. Methods in Enzymology, Goeddcl D.V. (cd.), 1985: 60-89.