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Bioluminescence by the squid Euprymna scolopes requires colonization of its light organ by the symbiotic luminous bacteriumVibrio fischeri . Investigation of the genetic determinants underlying bacterial symbiotic competence in this system has necessitated the continuing establishment and application of molecular genetic techniques in V. fischeri. We developed a procedure for the introduction of...
Our previous study demonstrated that a cis-acting element for neuron-specific splicing of human amyloid precursor protein (APP) gene existed in an intron region flanking to exon 8 (Yamada et al., 1993). In this study, we extended the analysis by deletion and mutagenesis experiments and identified two distinct cis-acting elements which modulate the splicing of exon 8 of the APP gene in a sequence-dependent...
The Aeromonas salmonicida A449 recA gene has been cloned, sequenced and expressed in vitro. The predicted amino acid sequence of A. salmonicida RecA was determined and, when compared to other RecA, was found to possess a number of domains identical to those characterized in Escherichia coli RecA. The A. salmonicida recA was mobilized into an E. coli recA mutant strain and was shown to allow increased...
An intronless gene encoding a putative G protein-coupled receptor was isolated from the genomic library of barnacle Balanus amphitrite Darwin, with probes obtained from degenerate polymerase chain reaction (PCR) primers used to amplify putative transmembrane regions. The cloned genome DNA specifies an open reading frame of 1431 bp encoding 476 amino acids with seven hydrophobic transmembrane (TM)-spanning...
Sequences representing the 2684 bp flanking the ipaH 4.5 gene on the invasion plasmid of Shigella flexneri 5 indicate an unusual fusion gene, designated ipgH, in which the first 27 amino acids (aa) are identical to ORF2 of IS629. The aa sequence encoded by the remainder of ipgH bears significant homology to Escherichia coli and to Salmonella typhimurium GlpT and UhpT proteins and...
A number of transcription factors that act as adaptor proteins have been found to contain an 87 amino acid domain called the bromodomain. In a study to identify and characterise bromodomain proteins expressed in chicken cells, a novel gene has been isolated which encodes five repeats of the bromodomain. In addition, the encoded protein, termed polybromo, contains four other domains: an unusual truncated...
We cloned the Aspergillus oryzae tannase gene using three oligodeoxyribonucleotide (oligo) probes synthesized according to the tannase N-terminal and an internal amino acid (aa) sequence. The nucleotide (nt) sequence of the tannase gene was determined and compared with that of a tannase DNA complementary to RNA (cDNA) by means of reverse transcriptase PCR. The results indicated that there was no...
Type 4 fimbriae are surface organelles produced by a wide range of bacterial pathogens. In Pseudomonas aeruginosa they are associated with a form of surface translocation known as twitching motility and have also been implicated as the receptor for a number of fimbrial-specific bacteriophages. The infrastructural machinery required for type 4 fimbrial biogenesis appears to be conserved as heterologous...
We report the cloning of a bromodeoxyuridine (BrdU)-sensitive transcript of 918 bp from an immortalized quail heart cell line containing an open reading frame (ORF) of 215 amino acids (aa) ( 23 kDa). Analysis of the secondary structure predicts two amphipathic α-helices with oppositely oriented amphipathic surfaces at the C-terminus of the protein. Each of the helices contains an LEA (late embryogenesis...
Transcription of genes from phage LL-H can be divided into an early phase and a late phase. The early gene region was located in a 5.9-kb segment of the phage LL-H genome and it was part of the sequence that completed the phage LL-H genome sequence, 34 659 bp in size. Phage LL-H is the first completely sequenced Lactobacillus phage. In the main coding strand of phage LL-H genome 48 putative ORFs...
A gene conferring resistance to fosmidomycin (Fs) was cloned from the gene pool of a wild-type strain of Escherichia coli. The cloned DNA fragment was sequenced and shown to encode a putative polypeptide of 406 amino acids (aa) with a molecular weight of 43 303. The gene mapped at 10.9 min on the E. coli chromosome and was designated fsr (fosmidomycinr esistance). Maxicell analysis revealed that...
Transfection of the mouse double minute 2 (Mdm2) oncogene has been found to induce immortalization of primary cells and to transform cultured cells. Amplification and/or overexpression of human MDM2 has been documented in a large percentage of human cancers. Mouse and human Mdm2 cDNA have been cloned from transformed cells and the cDNA sequence of both genes have been reported previously. In this...
Heterologous gene expression levels were measured in yeast using the Escherichia coli gusA gene (encoding β-d-glucuronidase) as a reporter. The influence of two major parameters, promoter activity and plasmid copy number, was studied. (1) Promoters used in this study ranged from the very weak constitutive KEX2, the regulated CYC1 and PGK and the mating type-specificMFα1 to the strong constitutive...
A new insertion sequence designated IS1394 was isolated from Pseudomonas alcaligenes NCIB 9867 (P25X) by entrapment in plasmid pUCD800 which carries the Bacillus subtilis sacB and sacR genes. The 1100-bp sequence contains 27-bp inverted repeats with 4 bp mismatch and has one long open reading frame, spanning 92.1% of the entire IS. The deduced 338 amino-acid sequence demonstrated homology (varying...
RT-PCR was used to examine the expression of LAR (encoding the leukocyte-common antigen-related protein tyrosine phosphatase) in normal human colon mucosa, and colon polyps and tumors. Although the LAR protein was not detected in the colon in a previous immunohistochemical study, amplification of a region of LAR between the most membrane proximal (eighth) fibronectin type-III (FN-III) repeat and...
Peripheral myelin protein 22 (PMP22) is expressed by Schwann cells in the peripheral nervous system (PNS), and mutations affecting the PMP22 gene are associated with hereditary motor and sensory neuropathies. We have previously defined the PMP22/EMP/MP20 gene family by characterizing the PMP22-related epithelial membrane protein-1 (EMP-1). We now report the identification of two additional members...
The mcr locus from Streptomyces lavendulae confers high level resistance (> 100 μg/ml) to mitomycin C (MC) and related mitomycins when cloned into Streptomyces lividans. Production of the mcrA gene product (MCRA) was shown to be MC-inducible by identification of MCRA (M r of 54 kDa) using Western blot analysis and enzyme linked immunosorbent assay (ELISA). The magnitude of MCRA production...
The asoB gene of Aeromonas salmonicida is located approximately 9 kb downstream of the structural gene (vap A) for the surface layer (A-layer). The nucleotide sequence of asoB was determined and found to encode a putative polytopic cytoplasmic membrane protein which exhibited homology to a number of bacterial transport proteins. Allele exchange mutagenesis of asoB resulted in a mutant (A449-D) which...
We have previously examined the efficiency of two Drosophila melanogaster promoters to enable reporter gene expression in embryos of the Australian sheep blowfly, Lucilia cuprina. Both the hsp70 heat-shock promoter and the actin5C promoter resulted in low levels of expression of a reporter gene in these embryos. In this study, the D. pseudoobscura hsp82 promoter (phsp82) was tested for its ability...
Recombinant retroviral (re-Rv) and adenoviral (re-Ad) vectors for delivery of two foreign genes were constructed, using the internal ribosomal entry site (IRES) of encephalomyocarditis virus (EMCV) which mediates initiation of cap-independent translation. The first gene encoded the hepatitis B surface antigen (HBsAg) and the second encoded human or murine B7-1 molecule, a cell surface protein which...
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