Cloning and expression of the two DNA fragments of the I-18 C region of Chironomus tentans. II. The effects of the applied technology

• Autorzy: Borowicz B P
• Języki publikacji: EN

Abstrakty

EN

The chromosomal I-18 C region of Chironomus tentans contains the I-18 C gene. Two different open reading frames (i.e. the ORF I and II) of two different transcripts (i.e. the 1.8 kb and 4.6 kb RNA) of the I-18 C gene of Chironomus tentans were isolated, at the DNA level, by the Polymerase Chain Reaction, then were cloned into the bluescript vector and finally were cloned into the pET-3a vector in order to translate them in T7 RNA polymerase / promoter system of E. coli. It was possible to obtain the ORF I overexpressed. In a case of the ORF II the low molecular weight polypeptide was detected, however it was not overexpressed, but still this polypeptide was strongly visible on the SDS-polyacrylamide gel.

PL

Region chromosomalny I-18 C Chironomus tentans zawiera gen I-18 C. Dwie odrębne otwarte ramy odczytu (tj. ORF I i II) genu I-18 C Chironomus tentans zostały wyizolowane, na poziomie DNA, przy użyciu Reakcji Łańcuchowej Polimerazy (PCR), a następnie zostały wklonowane do wektora blueskrypt i ostatecznie zostały wklonowane do wektora pET-3a w celu ich translacji w systemie promotora T7 RNA polimerazy w E. coli. Uzyskano bardzo dużą ekspresję ORF I. W przypadku ORF II wykryto obecność polipeptydu o niskiej masie cząsteczkowej i chociaż nie uzyskano jego bardzo dużej ekspresji, tym niemniej polipeptyd ten był silnie widoczny w żelu poliakryloamidowym z SDS.

Słowa kluczowe

EN

promoter system; cloning; I-18 C gene; T7 RNA polymerase; Chironomus tentans; polymerase chain reaction; DNA fragment

• Wydawca: -
• Czasopismo: Journal of Plant Protection Research
• Rocznik: 1998
• Tom: 38
• Numer: 1
• Opis fizyczny: p.42-58,fig.

Twórcy

autor

Borowicz B P

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

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