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The rrnA ribosomal RNA (rRNA) operon of Campylobacter jejuni (Cj) TGH9011 (ATCC43431) was cloned and sequenced to completion. rRNAs were then characterized by primer extension and S1 nuclease mapping analysis. The secondary structure models of Cj 16S and 23S rRNAs were constructed, and the models were compared to the corresponding models from other eubacterial rRNA. The analysis presented a typical...
The temperature-regulated SerH1 gene coding for an immunodominant surface glycoprotein (i-AgH1) of Tetrahymena thermophila has been sequenced. The gene is reproducibly rearranged during macronuclear development and steady state mRNA levels are present at < 36°C. The deduced i-Ag H1 amino acid (aa) sequence is rich in Ser, Thr and Cys, and contains three periods each consisting of 85 aa punctuated...
With the goal of understanding possible mechanisms of drug resistance by the protozoan parasite Entamoeba histolytica (Eh), two novel Eh P-glycoprotein (Pgp) genes (Eh pgp5 and Eh pgp6) were sequenced, and the expression of fourEh pgp genes determined in wild-type (wt) clone A and emetine-resistant (Em R ) clone C2 amebae. The Eh pgp5 gene encodes a 1301-amino acid (aa) protein that is similar...
Lactobacillus (Lb.) helveticus CNRZ32 possesses a 97-kDa metalloenzyme with aminopeptidase activity (PepN; EC 3.4.11.2). A 3.8-kb fragment encoding PepN was cloned into pIL253 and designated pSUW34. Transformation ofLactococcus (Lc.) lactis LM0230 with pSUW34 rersulted in >180-fold increase in general aminopeptidase (AP) activity using l-lysine-p-nitroanilide. Southern hybridization was conducted...
A 1.2-kb DNA fragment from Bacillus subtilis CD4 encoding endo-β-1,4-glucanase and cellobiase activities was cloned and expressed in Escherichia coli. Carboxymethylcellulase (CMCase) and cellobiase activities were detected in a cell-free lysate of recombinant (re-) E. coli (Ec). The re-enzymes were functional in Ec, as it utilized carboxymethylcellulose, soluble cellulose and cellobiose as sole...
The O antigens found in Salmonella enterica (Se) and Escherichia coli (Ec) show a great deal of diversity, and only three structures are known to be common to both genera. Two of them contain the 3,6-dideoxyhexose colitose, not found in other serogroups of the two species. The first of these is common to Ec O111 and Se O:35 (sv Adelaide); the other is found in both Ec O55 and Se O:50 (sv Greenside)...
A gene encoding ankyrin (Ank) was isolated from a genomic library of the plant pathogen Pseudomonas syringae pathovar syringae strain 61 (Pss61). The gene encodes an 183-amino-acid (aa) polypeptide which has homology to the 33-aa repeat domain of mammalian Ank and Ank homologs from other bacteria, animals and plants.
Mutations of genes involved in the STT1PKC1 pathway in yeast show staurosporine and temperature sensitivities (stt) which are suppressed by the addition of 1 M sorbitol [Yoshida et al., Mol. Gen. Genet. 242 (1994) 631-640]. Among the stt mutants, stt3-2 shares this phenotype. The STT3 gene encodes a novel 718-amino-acid protein with significant homology to potential transmembrane proteins of Caenorhabditis...
Based on the studies of the FliA protein in Bacillus subtilis (Bs) and Salmonella typhimurium (St), theEscherichia coli (Ec) fliA gene has been proposed to encode a flagellar-specific sigma factor, σ 28 . In this study, the complete nucleotide (nt) sequence of Ec fliA was determined. The fliA coding region consists of 717 nt starting with a GTG start codon and ending with a TAA stop...
An Escherichia coli strain overproducing the KpnI DNA methyltransferase (M KpnI) was constructed by cloning thekpnIM gene downstream from the inducible T7 phage 10 promoter. A method involving three chromatographic steps has been developed to purify M KpnI to homogeneity. The purified enzyme has a pH optimum around 7.3 and is inhibited by salts. M KpnI can be photolabeled by UV-irradiation...
Nocardia asteroides (Na) superoxide dismutase (SOD) has been implicated as a virulence factor that allows the organism to survive intracellular killing by phagocytic cells. A full-length Na sod gene from a pathogenic strain of Na (strain GUH-2) was cloned from a recombinant phage library using the Mycobacterium tuberculosis (Mt) sod gene (Mt sod) as a probe. The promoter region and structural gene...
The sequence surrounding the Azorhizobium caulinodans (Ac) regulatory nodD gene was analyzed. Upstream fromnodD and in the opposite orientation, two small open reading frames were identified (ORF1 and ORF2). The DNA sequence corresponding to ORF1, termed 1, is similar to a part of the insertion element IS51 from Pseudomonas savastanoi. Immediately downstream from nodD, a repeated element, δ1,...
Sequence similarity analysis has revealed that orf2, in the cell division 135-137° region of the Bacillus subtilis (Bs) chromosome, is the probable homolog of Escherichia coli murB (encoding a reductase involved in peptidoglycan synthesis). The amino-acid sequences of the two protein products show 24% identity (47% overall similarity), with several regions of higher similarity which may represent...
A 30-kDa protein accumulated upon induction by a high concentration of tyramine or dopamine in cells of Klebsiella aerogenes (Ka). These cells carried a plasmid (pAS123) that included the arylsulfatase operon (atsBA). Deletion analysis showed that the region essential for induction of the 30-kDa protein was located within a 2.0-kb cloned segment downstream of the atsBA operon. The nucleotide (nt)...
Previously, the double-transposon (Tn) mutant VAN20 of Vibrio anguillarum (Va) 775.17B was isolated. This mutant lacked a major surface antigen (MSA) suggested to be a lipopolysaccharide (LPS) and showed a 10 5 -fold increase in the 50% lethal dose (LD 50 ) when fish were infected intraperitoneally. In this study, the two Tn insertion sites within the chromosome were identified,...
We have cloned the Francisella tularensis (Ft) grpE-dnaK-dnaJ heat-shock genes which are organized in that order. These genes allow heterologous genetic complementation of each respective mutant strain of Escherichia coli (Ec) for bacteriophage λ growth. The nucleotide sequences of the Ft grpE-dnaK-dnaJ genes and the deduced amino-acid sequences share significant homologies with their respective...
We have constructed a new multipurpose stable shuttle vector for the fission yeast Schizosaccharomyces pombe (Sp). Plasmid pDblet was designed to provide convenient features for molecular work and to overcome the inconveniences of previously designed Sp vectors. It contains the Sp ura4 gene as selectable marker and a new highly efficient ARS (autonomously replicating sequence) element, allowing...
We have constructed a cloning vector based on plasmid mini-F for use in Escherichia coli. Plasmid pZC320 consists of the ori-2 replication unit of F that confers very low copy number (lcn), and includes the sop partition functions to insure stable plasmid maintenance in the absence of selection. A multiple cloning site (MCS) containing 16 unique restriction sites is located within the 5 end of...
A new enforcement (kyosei-) cloning plasmid vector, designated pKF4, was constructed which confers kanamycin resistance (Km R ) and enforces streptomycin sensitivity (Sm S ). Since it is important to employ restriction endonuclease (ENase) preparations free of exonuclease (Exo) activities for effective use of the kyosei-cloning procedure [Hashimoto-Gotoh et al., Gene 137 (1993) 211-216],...
Although drug-resistance markers have been used frequently for gene-disruption mutagenesis in Streptococcus pneumoniae, none has yet been shown to be free of dependence on local transcription for its expression. Indeed, the erythromycin-resistance marker (erm), originating in pAMβ1, has been used as an indicator of local transcription on several occasions. A procedure is demonstrated for evaluation...
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