Abstract
Transcription of the cellulosomal cellulase/hemicellulase genes of Clostridium cellulovorans has been investigated by Northern blot, reverse transcriptase PCR (RT-PCR), primer extension, and S1 nuclease analysis. Northern hybridizations revealed that the cellulosomal cbpA gene cluster is transcribed as polycistronic mRNAs of 8 and 12 kb. The 8-kb mRNA coded for cbpA and exgS, and the 12-kb mRNA coded for cbpA, exgS, engH, and engK. The sizes of the mRNAs were about 3 kb for engE, 1.8 kb for manA, 2.7 kb for xynA, and 4 kb for pelA, indicating monocistronic transcription of these genes. Primer extension and S1 nuclease analysis of C. cellulovorans RNA showed that the transcriptional start sites of cbpA, engE, manA, and hbpA were located 233, 97, 64, and 61 bp upstream from the first nucleotide of each of the respective translation initiation codons. Alignment of the cbpA, engE, manA, and hbpA promoter regions provided evidence for highly conserved sequences that exhibited strong similarity to the σA consensus promoter sequences of gram-positive bacteria.
| Original language | English |
|---|---|
| Pages (from-to) | 2520-2527 |
| Number of pages | 8 |
| Journal | Journal of Bacteriology |
| Volume | 185 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2003 Apr 1 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Immunology
Cite this
Transcription of Clostridium cellulovorans cellulosomal cellulase and hemicellulase genes. / Han, Sung Ok; Yukawa, Hideaki; Inui, Masayuki; Doi, Roy H.
In: Journal of Bacteriology, Vol. 185, No. 8, 01.04.2003, p. 2520-2527.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transcription of Clostridium cellulovorans cellulosomal cellulase and hemicellulase genes
AU - Han, Sung Ok
AU - Yukawa, Hideaki
AU - Inui, Masayuki
AU - Doi, Roy H.
PY - 2003/4/1
Y1 - 2003/4/1
N2 - Transcription of the cellulosomal cellulase/hemicellulase genes of Clostridium cellulovorans has been investigated by Northern blot, reverse transcriptase PCR (RT-PCR), primer extension, and S1 nuclease analysis. Northern hybridizations revealed that the cellulosomal cbpA gene cluster is transcribed as polycistronic mRNAs of 8 and 12 kb. The 8-kb mRNA coded for cbpA and exgS, and the 12-kb mRNA coded for cbpA, exgS, engH, and engK. The sizes of the mRNAs were about 3 kb for engE, 1.8 kb for manA, 2.7 kb for xynA, and 4 kb for pelA, indicating monocistronic transcription of these genes. Primer extension and S1 nuclease analysis of C. cellulovorans RNA showed that the transcriptional start sites of cbpA, engE, manA, and hbpA were located 233, 97, 64, and 61 bp upstream from the first nucleotide of each of the respective translation initiation codons. Alignment of the cbpA, engE, manA, and hbpA promoter regions provided evidence for highly conserved sequences that exhibited strong similarity to the σA consensus promoter sequences of gram-positive bacteria.
AB - Transcription of the cellulosomal cellulase/hemicellulase genes of Clostridium cellulovorans has been investigated by Northern blot, reverse transcriptase PCR (RT-PCR), primer extension, and S1 nuclease analysis. Northern hybridizations revealed that the cellulosomal cbpA gene cluster is transcribed as polycistronic mRNAs of 8 and 12 kb. The 8-kb mRNA coded for cbpA and exgS, and the 12-kb mRNA coded for cbpA, exgS, engH, and engK. The sizes of the mRNAs were about 3 kb for engE, 1.8 kb for manA, 2.7 kb for xynA, and 4 kb for pelA, indicating monocistronic transcription of these genes. Primer extension and S1 nuclease analysis of C. cellulovorans RNA showed that the transcriptional start sites of cbpA, engE, manA, and hbpA were located 233, 97, 64, and 61 bp upstream from the first nucleotide of each of the respective translation initiation codons. Alignment of the cbpA, engE, manA, and hbpA promoter regions provided evidence for highly conserved sequences that exhibited strong similarity to the σA consensus promoter sequences of gram-positive bacteria.
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UR - http://www.scopus.com/inward/citedby.url?scp=0344838397&partnerID=8YFLogxK
U2 - 10.1128/JB.185.8.2520-2527.2003
DO - 10.1128/JB.185.8.2520-2527.2003
M3 - Article
VL - 185
SP - 2520
EP - 2527
JO - Journal of Bacteriology
JF - Journal of Bacteriology
SN - 0021-9193
IS - 8
ER -