Efficiency of transcriptional terminators in Bacillus subtilis

Hess, Gerard F.; Graham, R. Scott

doi:10.1016/0378-1119(90)90425-q

Cited by 6 publications

(2 citation statements)

References 14 publications

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“…In the present study, we developed a similar system to produce tag‐fusing chimera polypeptides in B. subtilis using pWH1520, a shuttle vector for heterologous gene expression in B. megaterium [26]. The recognition sequence of factor Xa (IEGR), six successive histidine codons and a B. licheniformis β‐lactamase terminator sequence [27] with no termination codon in frame were designed to be aligned within the xylose isomerase gene under the xyl A promoter on pWH1520 (Fig. 2a ).…”

Section: Resultsmentioning

confidence: 99%

“…The plasmid pWH1520, a shuttle vector for heterologous gene expression in Bacillus megaterium [26], was used for expression from a truncated mRNA lacking a termination codon in B. subtilis . The recognition sequence of factor Xa (IEGR), six successive histidine codons and a Bacillus licheniformis β‐lactamase terminator sequence [27] with no termination codon in frame were designed to be aligned within the partial xylose isomerase gene under the xyl A promoter on pWH1520. This plasmid was then transformed into B. subtilis strain AMHG L1 (P spac‐ssrA ) or AMHG N1 (P spac‐ssrA ( DD )) [24].…”

Section: Methodsmentioning

confidence: 99%

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Trans‐translation mediated by Bacillus subtilis tmRNA

et al. 2002

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Trans‐translation, in which a ribosome switches between translation of an mRNA and a tmRNA, produces a chimera polypeptide of an N‐terminal truncated polypeptide and a C‐terminal tag‐peptide encoded by tmRNA. One of the tmRNA binding proteins, a ribosomal protein S1, has not been found in a group of Gram‐positive bacteria. In this study, the trans‐translation reaction with tmRNA from Bacillus subtilis belonging to this group was examined. When a truncated gene lacking a termination codon was expressed in B. subtilis, a 15‐amino acid tag‐peptide derived from tmRNA was identified in the C‐termini of the trans‐translation products. An identical tag‐peptide was also found at the C‐termini of the products from a truncated gene, when it was coexpressed with B. subtilis tmRNA in Escherichia coli. B. subtilis tmRNA was functional, although much less efficiently, in the in vitro poly(U)‐dependent tag‐peptide synthesis system of E. coli. A comparison of two bacterial tmRNAs suggests that the rule for determining the tag‐initiation point on tmRNA may be the same in Gram‐positive and Gram‐negative bacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Trans‐translation mediated by Bacillus subtilis tmRNA

et al. 2002

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Cloning sequencing and expression of the gene for cytochrome P450meg, the steroid-15β-monooxygenase from Bacillus megaterium ATCC 13368

Rauschenbach¹,

Isernhagen²,

Noeske-Jungblut³

et al. 1993

Molec. Gen. Genet.

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A 4.3 kb EcoRI fragment carrying the gene for cytochrome P450meg, the steroid-15 beta-monooxygenase from Bacillus megaterium ATCC 13368, was cloned and completely sequenced. The gene codes for a protein of 410 amino acids and was expressed in Escherichia coli and B. subtilis. Protein extracts from the recombinant E. coli strains were able to hydroxylate corticosteroids in the 15 beta position when supplemented with an extract from a P450- mutant of B. megaterium ATCC 13368 as a source of megaredoxin and megaredoxin reductase. In contrast, 15 beta-hydroxylation was obtained in vitro and in vivo without the addition of external electron transfer proteins, when cytochrome P450meg was produced in B. subtilis 168. Protein extracts from nonrecombinant B. subtilis 168 could also support the in vitro hydroxylation by cytochrome P450meg produced in E. coli.

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Chapter 6 Messenger RNA Decay and Maturation in Bacillus subtilis

Bechhofer

2009

Progress in Molecular Biology and Translational Science

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Efficiency of transcriptional terminators in Bacillus subtilis

Cited by 6 publications

References 14 publications

Trans‐translation mediated by Bacillus subtilis tmRNA

Trans‐translation mediated by Bacillus subtilis tmRNA

Cloning sequencing and expression of the gene for cytochrome P450meg, the steroid-15β-monooxygenase from Bacillus megaterium ATCC 13368

Chapter 6 Messenger RNA Decay and Maturation in Bacillus subtilis

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