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J Seijffers

Publications and source records attributed to J Seijffers.

8 recordsLinked to original sources
PubMed1990

Molecular characterization of genes coding for wild-type and sulfonylurea-resistant acetolactate synthase in the cyanobacterium Synechococcus PCC7942.

We present here the isolation and molecular characterization of acetolactate synthase (ALS) genes from the cyanobacterium Synechococcus PCC7942 which specify a sulfonylurea-sensitive enzyme and from the sulfonylurea-resistant mutant SM 3/20, which specify resistance to sulfonylurea herbicides. The ALS gene was cloned and mapped by complementation of an Escherichia coli ilv auxotroph that requires branched-chain amino acids for growth and lacks ALS activity. The cyanobacterial gene is efficiently expressed in this heterologous host. The ALS gene codes for 612 amino acids and shows high sequence homology (46%) at the amino acid level with ALS III of E. coli and with the tobacco ALS. The resistant phenotype is a consequence of proline to serine substitution in residue 115 of the deduced amino acid sequence. Functional expression of the mutant gene in wild-type Synechococcus and in E. coli confirmed that this amino-acid substitution is responsible for the resistance. Yet the deduced amino-acid sequence as compared with other ALS proteins supports the notion that the amino-acid context of the substitution is important for the resistance.

Acetolactate Synthase↗
PubMed1989

The 5'-flanking region of the gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase is crucial for growth of the cyanobacterium Synechococcus sp. strain PCC 7942 at the level of CO2 in air.

Transformation of the high-CO2-requiring mutants (hcr) O221 and E1 derived from the cyanobacterium Synechococcus sp. strain PCC 7942 by a wild-type DNA library restored their ability to grow at the level of CO2 in air. A plasmid (pE12) containing a 10-kilobase DNA insert was rescued from a O221 heterogenote and proved to transform both O221 and E1 to the wild-type phenotype. The capacity of the pE12 subclones to confer the wild-type phenotype to O221 transformants enabled the mapping of the mutation in O221 (designated hcrO221) within a 232-base-pair PstI-BstXI DNA restriction fragment. Sequence analysis revealed two open reading frames (ORFs) at positions -1745 to -1262 (ORFI) and -1218 to -393 (ORFII) upstream of the rbcL gene. A 3-kilobase PstI fragment of O221 was cloned, and hcrO221 was found to be a point mutation within the PstI-BstXI region -1309 nucleotides upstream of the rbcL gene. The significance of this flanking region for adaptation to air levels of CO2 was further demonstrated by the generation of new hcr mutants following insertional inactivation of wild-type DNA in the BstXI site. Electron microscopy revealed aberrant carboxysome structures in growing cells of the hcr mutants, a defect that was possibly related to the mutation, since transformation with pE12 derivatives restored the carboxysome structure to normal.

Air↗
PubMed1986

Controlled gene expression utilising lambda phage regulatory signals in a cyanobacterium host.

This study presents plasmid systems that utilize regulatory signals of bacteriophage Lambda to accomplish regulated expression of cloned genes in an A. nidulans R2 derivative strain. An operator-promoter region and the temperature-sensitive repressor gene cI857 of bacteriophage Lambda were employed. Linked to a cyanobacterial replicon, the plasmid vectors efficiently transformed Anacystis and were stably maintained within this host. The cat structural gene, encoding chloramphenicol acetyltransferase, was used to demonstrate that expression can be regulated by temperature shift. We have identified in extracts from the vector bearing Anacystis, a protein similar in size and immunology to the Lambda repressor. The systems described should allow controlled expression of adventitious genes in the cyanobacterial host.

Acetyltransferases↗
PubMed1977

Osmoregulation in symbiosis-independent mutants of Bdellovibrio bacteriovorus.

Bdellovibrios capable of axenic growth grow in a cell-free medium at a rate considerably lower than that attainable in a two-membered culture with Escherichia coli. The axenic growth rate may be improved either by adjustment of the osmosity of the medium or by the addition of low concentrations of spermine.

Bdellovibrio↗
PubMed1976

Effect of virginiamycin on the growth cycle of Bdellovibrio.

The two components of virginiamycin, virginiamycin M (VM) and virginiamycin S (VS), were used to explore the life cycle of symbiosis-dependent and -independent strains of Bdellovibrio bacteriovorus during multiplication in a two-membered system with either living or heat-inactivated Escherichia coli or in axenic cultures. Relatively high concentrations of these inhibitors separately were required to stop growth under all the conditions, but the minimum inhibitory concentration of the single components was reduced 1,000-fold by the association of VM and VS. No dissociation between mass growth and cell division was observed with VM; VS specifically halted cell division without affecting the kinetics of macromolecules formation and overall growth. This effect on cell division was only obtained when the antibiotic was added during the first half of the multiplication cycle and was reversible at any time.

Bacteria↗
PubMed1975

Symbiosis-independent and symbiosis-incompetent mutants of Bdellovibrio bacteriovorus 109J.

Symbiosis-independent (Sin) mutants were isolated from the symbiosis-dependent and symbiosis-competent (Sdcomp+) Bdellovibrio bacteriovorus 109J. Independently isolated Sin mutants were examined for their symbiosis competence and most were found to be comp+. Bdellovibrios comp- were selected from the Sincomp+ mutants. The Sincomp+ bdellovibrios are always at a selective disadvantage, either against Sincomp- bdellovibrios (in organic medium) or against Sdcomp+ bdellovibrios (in buffer with Escherichia coli cells).

Bacteria↗
PubMedDate not supplied

A new hybrid plasmid capable of transforming Escherichia coli and Anacystis nidulans.

We have constructed a hybrid plasmid, pDF30, by combining the 8-kb pDF3 plasmid derived from the cyanobacterium Anacystis nidulans 6311 with the Escherichia coli vector pBR325. pDF30 transforms, replicates and confers chloramphenicol resistance (Cmr) and ampicillin resistance (Apr) on both A. nidulans and E. coli. The level of resistance to ampicillin in A. nidulans transformants, although low, is above background resistance and an Apr activity was demonstrated in cell-free extracts of A. nidulans that harbored pDF30. pDF30 is stably maintained within E. coli and can be isolated intact from A. nidulans after several months of subculture under chloramphenicol selection and by this criterion is stable in vivo. The unique restriction sites for XhoI, SalI and EcoRI can be used for molecular cloning of chromosomal genes in E. coli and A. nidulans. Until now pDF3 is phenotypically cryptic in both A. nidulans and E. coli. Examination of the polypeptides encoded on and expressed by pDF30 in E. coli minicells revealed only gene products from pBR325 but none from pDF3.

Base Sequence↗