Gene
The gene organization has been determined for an 8-kb portion of the Streptomyces coelicolor chromosome close to the origin of DNA replication (oriC). Hybridization and DNA sequence analyses revealed the presence of five open reading frames (ORFs) oriented in the same direction as the proximal dnaA and dnaN genes. The deduced products of three of the ORFs have been identified as the S. coelicolor...
A 4906-bp DNA fragment, which complemented Schizosaccharomyces pombe strains partly defective in pyruvate dehydrogenase (PDH), was cloned and sequenced. The fragment contained an open reading frame (ORF) of 366 amino acids (aa), which showed 62% identity with the Saccharomyces cerevisiae gene encoding the PDH E1β subunit, and significant similarity to subunits from a number of other 2-oxo-acid dehydrogenase...
The amitochondrial (a-mt) protozoa include four groups of organisms that are of interest as important human parasites and as probable descendents of the earliest branches of eukaryotic evolution. These organisms have not been directly compared in terms of structure and function of a specific molecule. We sequenced portions of their rRNA-encoding genes coding for the internal transcribed spacers...
Promoter analysis was performed on the Rhizopus niveus 3-phosphoglycerate kinase 2-encoding gene (pgk2), one of the two pgk genes (pgk1 and pgk2) from this filamentous fungus sequenced so far. Deletion mutants of the promoter region were fused to the Escherichia coli uidA gene (which codes for β-glucuronidase; GUS), and introduced into R. niveus to measure the intracellular GUS activities of the...
The nucleotide sequence of the Rhizobium meliloti (Rm) fliP gene was determined. Rm strains carrying insertions within this gene were non-motile, lacked flagella and formed normal N 2 -fixing root nodules on alfalfa. The FliP protein showed similarity to several bacterial gene products involved in pathogenicity in both plant and animal pathogens. It is likely that all of these proteins...
Production of the plasmid-pXO2-encoded capsule by Bacillus anthracis is required for full virulence of the organism. The induction of capsule synthesis in vitro requires growth in the presence of bicarbonate and CO 2 ; however, little else is known about the regulation of capsule synthesis and the role it plays in the expression of virulence. Recently, transposon Tn917 mutagenesis of B....
The gene vapT, encoding VapT, one of the extracellular sodium dodecyl sulfate (SDS)-resistant alkaline serine proteases (Serp) from the Gram - Vibrio metschnikovii strain RH530 has been cloned in Escherichia coli. The recombinantE. coli produced a protease which co-migrated with VapT on gelatin polyacrylamide gels. The nucleotide (nt) sequence of the cloned vapT revealed a single open reading...
The segment of the rho gene that harbors the ts15 mutation was isolated and sequenced. The mutation, an IS1 insertion, changed the sequence of the C-terminal nine amino-acid residues, altering seven and eliminating the last two.
The DNA encoding the subunits of the ATP synthase (F 1 F o ) of Streptomyces lividans 66 strain 1326 was identified using oligodeoxyribonucleotide probes derived from the N-terminal sequence of subunit γ of the F 1 complex. The complete nucleotide sequence of the operon was determined. The atp operon contains nine genes, atpIBEFHAGDC, encoding the eight structural components...
In order to characterize individual protein components of the mitochondrial (mt) ribosome for regulatory, functional and evolutionary studies, the yeast nuclear gene MRP-L4 (accession No. Z30582), coding for the mt ribosomal protein (MRP) YmL4, has been cloned using oligodeoxyribonucleotides (oligos) deduced from a partial amino acid (aa) sequence [Graack et al., FEBS Lett. 242 (1988) 4-8] as screening...
The nucleotide sequence of the recA gene of Vibrio cholerae (Vc) has been determined. The amino acid (aa) sequence of the protein product is very similar to other known RecA aa sequences. However, this sequence does not agree with a previously reported Vc RecA aa sequence [Ghosh et al., Nucleic Acids Res. 20 (1992) 372].
We have analyzed the compositional distributions of coding sequences and their different codon positions, as well as the codon usage of the nuclear genes of Plasmodium falciparum, a parasite characterized by an extremely GC-poor genome. As expected, coding sequences are AT-rich, codon usage is strongly biased towards A or T in third codon positions, and some particular amino acids (aa) are especially...
A 6-kb EcoRI genomic DNA fragment of Coxiella burnetii, isolated from a recombinant bacteriophage λZapII library, allowed heterologous genetic complementation of Escherichia coli deleted for its dnaJ gene. The C. burnetti dnaJ gene was expressed in E. coli and identified by Western blot analysis using polyclonal antibodies raised against purified E. coli DnaJ protein. Deletion mapping and genetic...
The cloning and sequence of the 5 untranslated region (5 -UTR) of the Bacillus licheniformis (Bl) 6816 subtilisin Carlsberg gene (subC) are reported here. The 5 and 3 ends of subC transcripts were characterized, and the promoter identified. Expression was studied using a fused lacZ reporter gene integrated into the chromosome of heterologous host Bacillus subtilis (Bs). βGal activities of...
A Saccharomyces cerevisiae GAL7 expression vector for the production of protein fusions to glutathione S-transferase (GST) has been constructed. Using this vector, a GST fusion to human papillomavirus type 6 (HPV-6) E7 protein was produced and purified by affinity chromatography in a single step, at a yield of 2 μg/ml of culture. The E7 portion of the fusion protein was phosphorylated, in contrast...
The B-subunit pentamer(s) (EtxBp) of Escherichia coli heat-labile enterotoxin (LT) are secreted from Vibrio cholerae via the general secretion pathway (GSP), but remain periplasmic in E. coli. In order to determine if other Gram - bacteria were also able to secrete the ExtBp, the etxB gene, which encodes EtxB was introduced into different bacteria. Of the bacteria examined, most species...
Degenerate oligodeoxyribonucleotides complementary to sequences encoding conserved amino acid (aa) motifs ind -alanine:d-alanine ligases (Ddl) were used to amplify approx. 600-bp fragments from glycopeptide-resistant strains ofLeuconostoc mesenteroides (Lm), Lactobacillus plantarum, La. salivarius and La. confusus, and from a susceptible strain of La. leichmannii. Comparison of the deduced aa sequences...
The Escherichia coli (Ec) torC AD operon encoding the trimethyl amine N-oxide (TMAO) reductase system is induced by both TMAO and anaerobiosis. The tor regulatory regions from bacteria related to Ec have been amplified by the polymerase chain reaction (PCR) using degenerate oligodeoxyribonucleotide primers based on conserved sequences of the tor products. The amplified regions from Salmonella enteritidis...
Plasmids pLTV1 and pHV33, capable of replicating in both Gram + and Gram - bacterial hosts (shuttle vectors), when derived from the Escherichia coli strain HB101, were inactive in an electro-transformation assay employing theBacillus anthracis strains ΔAmes-1 and ΔV1B-1 as recipients. The same plasmids isolated from the DNA methyltransferase (MTase)-deficient E. coli strain GM2929...
A library of Actinomyces naeslundii T14V DNA was constructed in plasmid pUC18 and from this several sucrose-positive clones were isolated. Evidence was obtained that all these clones contained the same gene. One clone, which carried a plasmid that was named pPNG102, was chosen for further study. It was found that the enzyme specified by this plasmid hydrolyzed sucrose, raffinose, inulin and levan,...