Cloning of the DNA fragments reflecting the open reading frame I and II of the I-18 C gene of Chironomus tentans. V. Identification of the bacterial colonies, containing the recombinant bluescript plasmids

• Autorzy: Borowicz B P
• Języki publikacji: EN

Abstrakty

EN

The technology applied for the cloning of the open reading frame (ORF) I and II reflecting DNA fragments in regard to the identification of the bacterial colonies, of E. coli XL-1 strain, containing the recombinant bluescript plasmids, is described. The particular steps include: the α-complementation test, growing of the preliminary selected bacterial colonies in culture, isolation of the plasmids from these colonies, the Nde I and Bam HI digestion of the plamids mini-preparations, analysis of the digestion products and the identification of the bacterial colonies, containing the bluescript plasmids with the cloned inserts by the agarose gel electrophoresis (2% gel, 1 x TBE buffer).

PL

Opisano technologię zastosowaną do klonowania fragmentów DNA odzwierciedlających otwartą ramę odczytu I i II genu I-18 C Chironomus tentans, w zakresie: identyfikacji kolonii bakteryjnych szczepu XL-1, zawierających zrekombinowane plazmidy blueskrypt. Poszczególne etapy obejmowały: test α-komplementacji, hodowlę wstępnie wybranych kolonii bakteryjnych, izolację plazmidów bakteryjnych poszczególnych kolonii z tych hodowli, trawienie mini-izolatów plazmidów Nde I i Bam HI, analizę produktów trawienia i identyfikację kolonii bakteryjnych, zawierających plazmidy blueskrypt z wklonowanymi wstawkami przy zastosowaniu elektroforezy w żelu agarozowym (2% żel, 1 x stężony bufor TBE).

Słowa kluczowe

EN

pET-3a vector; promoter system; cloning; I-18 C gene; T7 RNA polymerase; plasmid; Chironomus tentans; polymerase chain reaction; DNA fragment; identification

• Wydawca: -
• Czasopismo: Journal of Plant Protection Research
• Rocznik: 1999
• Tom: 39
• Numer: 1
• Opis fizyczny: p.59-62

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 Reaction. 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 Rcaction. 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 (in English). 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 (in English). 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 (in English). 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 technologics (in English). 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 (in English). J. Plant Protection Res., 37 (1/2): 165-171.
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