Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays

Wendisch VF, Zimmer DP, Khodursky A, Peter B, Cozzarelli N, Kustu S (2001)
Analytical Biochemistry 290(2): 205-213.

Zeitschriftenaufsatz | Veröffentlicht| Englisch
 
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Autor/in
Wendisch, Volker F.UniBi ; Zimmer, D. P.; Khodursky, A.; Peter, B.; Cozzarelli, N.; Kustu, S.
Abstract / Bemerkung
Bacterial messenger RNA (mRNA) is not coherently polyadenylated, whereas mRNA of Eukarya can be separated from stable RNAs by virtue of polyadenylated 3 ' -termini, We have developed a method to isolate Escherichia coil mRNA by polyadenylating it in crude cell extracts with E. coil poly(A) polymerase I and purifying it by oligo(dT) chromatography. Differences in lacZRNA levels were similar with purified mRNA and total RNA in dot blot hydridizations for cultures grown with or without gratuitous induction of the lactose operon, More broadly, changes in gene expression upon induction were similar when cDNAs primed from mRNA or total RNA with random hexanucleotides were hydridized to DNA microarrays for the E. coil genome. Comparable signal intensities were obtained with only 1% as much oligo(dT)-purified mRNA as total RNA, and hence in vitro poly(A) tailing appears to be selective for mRNA. These and additional studies of genome-wide expression with DNA microarrays provide evidence that in vitro poly(A) tailing works universally for E. coil mRNAs, (C) 2001 Academic Press.
Stichworte
polyadenylation; genome; yeast; sequence; lactose operon; messenger-rna; gene-expression; poly(a) polymerase; polyadenylation; messenger rna; DNA microarrays; escherichia coli; polymerase; bacteria; protein
Erscheinungsjahr
2001
Zeitschriftentitel
Analytical Biochemistry
Band
290
Ausgabe
2
Seite(n)
205-213
ISSN
0003-2697
Page URI
https://pub.uni-bielefeld.de/record/1895392

Zitieren

Wendisch VF, Zimmer DP, Khodursky A, Peter B, Cozzarelli N, Kustu S. Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Analytical Biochemistry. 2001;290(2):205-213.
Wendisch, V. F., Zimmer, D. P., Khodursky, A., Peter, B., Cozzarelli, N., & Kustu, S. (2001). Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Analytical Biochemistry, 290(2), 205-213. doi:10.1006/abio.2000.4982
Wendisch, V. F., Zimmer, D. P., Khodursky, A., Peter, B., Cozzarelli, N., and Kustu, S. (2001). Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Analytical Biochemistry 290, 205-213.
Wendisch, V.F., et al., 2001. Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Analytical Biochemistry, 290(2), p 205-213.
V.F. Wendisch, et al., “Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays”, Analytical Biochemistry, vol. 290, 2001, pp. 205-213.
Wendisch, V.F., Zimmer, D.P., Khodursky, A., Peter, B., Cozzarelli, N., Kustu, S.: Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Analytical Biochemistry. 290, 205-213 (2001).
Wendisch, Volker F., Zimmer, D. P., Khodursky, A., Peter, B., Cozzarelli, N., and Kustu, S. “Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays”. Analytical Biochemistry 290.2 (2001): 205-213.

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