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Two regions from the genome of the virulent Lactobacillus delbrueckii subsp. lactis bacteriophage LL-H were sequenced (2330 and 12939 bp; 44% of the 34.6-kb genome). Together with the previously sequenced region containing the major capsid protein-encoding gene (2498 bp), the sequence had 21 open reading frames (ORFs) on the main coding strand. Only two putative ORFs were detected on the complementary...
CDK2 (cyclin-dependent kinase 2) is a serine/threonine kinase which is involved in regulating S-phase entry in higher eukaryotes. To investigate the transcriptional control of this gene, a 13-kb Xenopus laevis genomic clone containing the 5 flanking sequences was isolated. A 2.7-kb fragment containing the promoter region was sequenced and the transcription start point (tsp) was determined by primer...
Previous studies have indicated that there may be uncleavable TaqI sites (TCGA) within the mouse myogenic gene,Myo-D1 . Fragments of DNA bearing most of the presumed insensitive TaqI sites have been reproduced using PCR. The presence of each of the originally uncleavable TaqI sites has been confirmed and each TaqI site has been shown to be sensitive to TaqI hydrolysis in PCR-synthesised genomic DNA...
Deletions in either of the genes in the strA-strB gene pair of Erwinia amylovora plasmid pEa34 resulted in a dramatic decrease in streptomycin resistance (Sm R ), but Sm R was restored to high levels by complementation. When strA andstrB were cloned separately on a lacI q /Ptac-based expression vector in Escherichia coli, only the protein encoded bystrA was produced. When a...
Temporal and spatial expression of muscle actin (MA)-encoding genes in the Halocynthia roretzi MA (HrMA) cluster was examined by whole-mount in situ hybridization and Northern blot analyses. Expression of these MA was restricted to larval muscle cells. None of the MA from the cluster was expressed in any adult tissues examined, including two different types of adult muscle tissue, body-wall muscle...
Full-length, sense-strand, long interspersed element-1 (LINE-1 or L1) RNA is found as an RNA-protein complex in mouse embryonal carcinoma cells. Since this complex is a likely intermediate in LINE-1 transposition, its RNA may be enriched for the functional, or active, subset of mouse L1 sequences. For this reason, a cDNA library was constructed from RNA prepared from these ribonucleoprotein particles...
A cDNA encoding ribosomal (r) protein L18 of Arabidopsis thaliana (At) was isolated and characterized. The nucleotide sequence contains a 563-bp open reading frame that encodes a 20.9-kDa basic protein. Amino-acid comparison indicated that the predicted L18 r-protein of At has a high degree of homology with L18 of distantly related organisms such as yeast, Xenopus laevis and rat. Genomic DNA analysis...
We have characterized the immediate (465 bp) 5 -flanking region of the rat phenylalanine hydroxylase (PAH)-encoding gene. This sequence shows considerable similarity to the 5 -flanking region of the human PAH gene [Konecki et al., Biochemistry 31 (1992) 8363-8368]. Both sequences lack obvious TATA elements; however, putative regulatory sites, including a potential cyclic AMP-response element and...
We report the isolation and characterization of a chicken cDNA which putatively encodes an actin-like protein (chACTL). This 394-amino-acid (aa) polypeptide shares sequence homology (81, 70 and 67% identical aa, respectively) with three actin-related proteins (ARP) described for Drosophila melanogaster (ARP14D), Caenorhabditis elegans (ACTL) and Saccharomyces cerevisiae (ACT2). At least six chACTL...
The human poliovirus (PV) receptor (PVR) is a member of the immunoglobulin (Ig) superfamily with unknown cellular function. We have isolated a human PVR-related (PRR) cDNA. The deduced amino acid (aa) sequence of PRR showed, in the extracellular region, 51.7 and 54.3% similarity with human PVR and with the murine PVR homolog, respectively. The cDNA coding sequence is 1.6-kb long and encodes a deduced...
The pLEX series of vectors was developed for the stable expression of exogenous genes in the protozoan parasiteLeishmania . These pUC-based constructs contain one of three independent selectable markers and a multiple cloning site inserted between the upstream and downstream untranslated regions of the previously cloned Leishmania major HEXBP gene. Selection was based on resistance to the aminoglycosides,...
The overproduction of P-glycoprotein (Pgp) has been associated with the development and maintenance of the multidrug resistant (MDR) phenotype, although the regulatory events responsible have not yet been elucidated. We have analyzed the overexpression of the TATA-less hamster class-I Pgp-encoding gene (Pgp1) in several MDR Chinese hamster cell lines. The MDR lung cell line DC-3F/VCRd5L, as well...
The bamHIC gene, controlling the BamHI restriction-modification (R-M) system can functionally be replaced by providing pvuIIC or smaIC in trans. C BamHI, the protein product encoded by bamHIC, has been purified and shown to bind a 345-bp DNA fragment within the BamHI R-M system.
An individual strain of Neisseria gonorrhoeae may produce up to 16 different DNA methyltransferases (MTases). We have used a novel cloning system that is able to detect MTase clones in the absence of direct selection [Piekarowicz et al., Nucleic Acids Res. 19 (1991)1831-1835] to identify 14 different MTase clones. Initial characterization of these clones indicates that at least seven of these MTases...
The DNA methylation pattern of the promoter (pNF-L) region of the rat light-neurofilament-encoding gene (NF-L), a neuron-specific gene, was assessed in NF-L expressing and non-expressing cell lines and tissues by genomic sequencing using PCR amplification of bisulfite-modified DNA. We analysed twenty-five potential CpG methylation sites between nucleotide (nt) positions -311 and +103 of pNF-L, containing...
The structural gene (dyrA) encoding dihydrofolate reductase (DHFR) of Thermotoga maritima has been cloned, sequenced and expressed in Escherichia coli. The dyrA gene, located immediately upstream from the gene encoding aspartate carbamoyltransferase (pyrB), encodes a highly thermostable enzyme with a distinct thermophilic activity profile. Important structural features are conserved among all bacterial...
Small G-proteins encoded by the ras-like ypt genes are ubiquitous in eukaryotic cells. They have been shown to play an essential role in membrane vesicle transport. We have isolated four ypt genes, yptC1, yptC4, yptC5 and yptC6, fromChlamydomonas reinhardtii (Cr) genomic and cDNA libraries. Three of them, yptC1, yptC4 and yptC5, are close homologues of ypt genes previously found in the multicellular...
Lipases, serine esterase enzymes, play an essential role in the mycoplasmal nutritional requirement for long-chain fatty acids. Although the lipase(s) activity in different mycoplasma species has been investigated, the molecular biology of the corresponding genes has not been studied. Using a single-primer PCR technique combined to more classical cloning systems, an operon containing three open reading...
The ability of macrophages to kill some kinds of tumor cells is dependent upon the production of the free radical nitric oxide (NO) by the inducible enzyme NO synthase (iNOS; EC 1.14.13.39). Expression of the iNOS gene is induced by lipopolysaccharide (LPS) and augmented by interferon-γ (IFN-γ). Two regions of the iNOS promoter are known to regulate induction, a promoter proximal region I (RI) and...
Cytochrome P450 2B1, encoded by CYP2B1, and cytochrome P450 2B2, encoded by CYP2B2, are inducible in rat liver by phenobarbital (PB). We have used cultured adult rat hepatocytes to study molecular mechanisms regulatingCYP2B1CYP2B2 transcription. Northern blot analysis demonstrated that the endogeneousCYP2B1CYP2B2 genes were inducible by PB in such cultures. A PB-responsive element (PBRE) conferring...
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