Synthesis and secretion of an Erwinia chrysanthemi pectate lyase in Saccharomyces cerevisiae regulated by different combinations of bacterial and yeast promoter and signal sequences

Laing, Elizna; Pretorius, Isak S.

doi:10.1016/0378-1119(92)90159-m

Cited by 32 publications

(23 citation statements)

References 46 publications

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“…Total RNA was isolated by an adaptation of the protocol of Laing & Pretorius (1992). An overnight E. coli DH5a culture was diluted into pre-warmed Luria-Bertani broth, with antibiotic selection as required, and grown to an OD 600 of 0?7-0?8 at 37 uC, with constant shaking.…”

Section: Methodsmentioning

confidence: 99%

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

Gardner

Rawlings

2004

Microbiology

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Two closely related but compatible plasmids of the IncQ-2a and IncQ-2b groups, pTF-FC2 and pTC-F14, were discovered in two acidiphilic chemolithotrophic bacteria. Cross-complementation and cross-regulation experiments by the replication proteins were carried out to discover what changes were necessary when the plasmids evolved to produce two incompatibility groups. The requirement of a pTC-F14 oriV for a RepC DNA-binding protein was plasmid specific, whereas the requirement for the RepA helicase and RepB primase was less specific and could be complemented by the IncQ-2a plasmid pTC-FC2, and the IncQ-1b plasmid pIE1108. None of the IncQ-1a plasmid replication proteins could complement the pTC-F14 oriV, and pTC-F14 and RSF1010 were incompatible. This incompatibility was associated with the RepC replication protein and was not due to iteron incompatibility. Replication of pTC-F14 took place from a 5?7 kb transcript that originated upstream of the mobB gene located within the region required for mobilization. A pTC-F14 mobB-lacZ fusion was regulated by the pTC-F14 repB gene product and was plasmid specific, as it was not regulated by the RepB proteins of pTF-FC2 or the IncQ-1a and IncQ-1b plasmids. Plasmid pTC-F14 appears to have evolved independently functioning iterons and a plasmid-specific RepC-binding protein; it also has a major replication transcript that is independently regulated from that of pTF-FC2. However, the RepA and RepB proteins have the ability to function with either replicon.

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Section: Methodsmentioning

confidence: 99%

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

Gardner

Rawlings

2004

Microbiology

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“…Isolation of total mRNA. The yeast total RNA isolation protocol of Liang & Pretorius (1992) ACYC, which contains the complete ars operon from At. ferrooxidans.…”

Section: Methodsmentioning

confidence: 99%

The divergent chromosomal ars operon of Acidithiobacillus ferrooxidans is regulated by an atypical ArsR protein

Butcher

Rawlings

2002

Microbiology

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The chromosomal arsenic-resistance (ars) operon of Acidithiobacillus ferrooxidans is atypical in that it is divergent, with its arsCR and arsBH genes transcribed in opposite directions. Furthermore, the amino-acid sequence of the putative ArsR-like regulator of the ars operon is not conserved in regions that have been shown to be responsible for binding to arsenic. Instead, the ArsR-like protein of At. ferrooxidans is related to a group of unstudied ArsRlike proteins that have been found to be associated with chromosomal ars-like operons identified during genome-sequencing projects. Using arsB-lacZ, arsR-lacZ and arsC-lacZ fusions, it was shown that the ArsR-like protein of At. ferrooxidans is a repressor of the arsBH and arsRC genes of this organism, and that induction of gene expression took place when either AsIII (arsenite) or AsV (arsenate) were added. Deletion of 19 aa from the C terminus of the 118 aa ArsR protein did not affect the regulation of its activity, but deletion of an additional 28 aa inactivated ArsR. Northern-blot hybridization suggested that on induction of expression, the arsRC genes were transcribed in greater quantities than the arsBH genes, but that the level of induction was not affected by the form of arsenic added (AsIII or AsV).

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“…YCplaclIIbas&l plasmid pPR4 (Ap ~ LEU2 ADHle-MFals-celA-TRP5r) was maintained as autonomous circular minieh/omosomes in the transformed yeast cells. The mitotic stability of YCplaclII and pPR4 was determined as described by Laing and Pretorius (1992).…”

Section: Microbial Strains Plasmids and Growth Conditionsmentioning

confidence: 99%

“…coli DH5ct were used (Sambrook et al, 1989). DNA amplification by the polymerase chain reaction (PCR) technique, RNA isolation from yeast cells, Southern blot hybridisation and Northern blot hybridisation were performed as described by Laing and Pretorius (1992) and Sambrook et al (1989). Computer-aided analysis was done using the NCBI Blast E-mail Service (Altsehul et al, 1990).…”

Section: Microbial Strains Plasmids and Growth Conditionsmentioning

confidence: 99%

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Expression of the Ruminococcus flavefaciens cellodextrinase gene in Saccharomyces cerevisiae

1995

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Synthesis and secretion of an Erwinia chrysanthemi pectate lyase in Saccharomyces cerevisiae regulated by different combinations of bacterial and yeast promoter and signal sequences

Cited by 32 publications

References 46 publications

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

The divergent chromosomal ars operon of Acidithiobacillus ferrooxidans is regulated by an atypical ArsR protein

Expression of the Ruminococcus flavefaciens cellodextrinase gene in Saccharomyces cerevisiae

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