Gene
The cbbL and cbbS genes, encoding ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), were cloned and sequenced from a thermophilic hydrogen-oxidizing bacterium, Pseudomonas hydrogenothermophila strain TH-1. ThecbbL gene encoded a 474-amino-acid (aa) protein (53 285 Da); cbbS encoded a 124-aa protein (14 656 Da). An ORF found downstream from the cbbLS genes encoded a 267-aa protein (29 565...
Regulation of competence for genetic transformation in Streptococcus pneumoniae involves the comAB locus and a small extracellular protein, the competence factor (CF). The comA or comB mutations block both spontaneous competence induction and elaboration of CF, yet permit induction of competence by added CF and subsequent transformation at normal levels. Sequence and genetic studies showed that the...
A series of general-purpose plasmid vectors based on the phage λ origin of replication (ori) has been constructed. Each vector consists of a backbone plasmid encoding chloramphenicol resistance (Cm R ) and containing a unique HaeII site into which the lacZα-complementing multiple cloning site (MCS) region of an established vector was inserted. To increase the cloning potential of the inserted...
The gene encoding the RNA-dependent protein kinase (PKR) was cloned from mouse genomic DNA and characterized by restriction mapping, Southern blot analysis and sequencing. The Southern analyses were consistent with the presence of a single copy of the Pkr gene in the mouse haploid genome. The genomic nucleotide (nt) sequence, when compared to that of previously determined cDNA nt sequences, revealed...
A 2-kb 5 fragment of hPTN (human pleiotrophin-encoding) genomic DNA was sequenced. Within this region we identified a new, upstream exon (U2). Cloning and sequencing of the PCR products of cDNAs from a human melanoma cell line revealed that a 401-bp intron was spliced out between exon U2 and a previously described untranslated exon U1. Our analysis also revealed that previously reported transcription...
In order to identify specific DNA sequences useful as genetic landmarks in the construction of a complete map of the human mdr1 (multidrug-resistance) gene, we investigated the introns in the central region. In intron 14, we identified a long stretch of a homopyrimidine homopurine sequence most probably adopting an unconventional DNA conformation, followed by a cluster of three Alu repeated...
The human PITSLRE protein kinases (PK), members of the p34 c d c 2 kinase family named according to the single amino acid (aa) code of an important (PSTAIRE) regulatory region [Meyerson et al., EMBO J. 11 (1992) 2909-2917], are candidate tumor suppressor gene(s) localized to human chromosome 1p36.2 and a syntenic region of mouse chromosome 4 [Lahti et al., Nature Genet. 7 (1994)...
We report the isolation and sequencing of MTJ1, a 1792-bp cDNA from an M27 murine lung carcinoma cell line. The largest ORF within MTJ1 encodes a 63869-Da protein, containing a 73-amino-acid (aa) sequence (the J domain) that is conserved in proteins of the DnaJ family of chaperonins. The J domain of MTJ1 is bracketed by potential transmembrane domains in a similar configuration to the J domain of...
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...
The genomic region encoding the core (C) protein (amino acids 1-162) of hepatitis C virus (HCV) was expressed inEscherichia coli as a recombinant (re-) protein with the maltose-binding protein (MBP) using the prokaryotic expression vector pMAL-CR1. The fusion protein (C MBP) was identified as a approx. 62-kDa polypeptide by immunoblot analysis using antiserum to MBP and HCV-infected human sera....
A genomic DNA encoding a mouse adrenocorticotropin (ACTH) receptor was isolated. The predicted 296-amino-acid sequence showed 88.9 and 78.7% identities to the human and bovine homologues, respectively.
The murine manganous superoxide dismutase-encoding gene (MnSOD) is highly homologous in both sequence and organization to its rat homolog. MnSOD is encoded by five exons spanning approx. 7 kb of DNA. RNA blot analysis indicated multiple RNA species, with the major RNA corresponding to a 960-bp message. This major transcript is highly inducible by tumor necrosis factor (TNF) in murine fibroblasts...
To investigate the structure and regulation of the genes encoding components of the basal RNA polymerase II transcriptional machinery, the Drosophila melanogaster genes encoding transcription factor IIB (TFIIB) and the TATA-box-binding protein (TBP) were isolated and characterized. In the TBP gene, a single intron bisects the sequences that encode the two repeated DNA-binding domains of TBP, which...
Many eukaryotic DNA sequences, especially lenti-retrovirus proviral genomes and their env genes, are unstable when cloned in high-copy-number plasmids in Escherichia coli. Stability can be increased by the use of low-copy-number vectors, although plasmid yields are low. Vectors are described here that contain the intermediate-copy-number P15Aori for cloning, stable propagation and higher-yield production...
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...
We have isolated, based on reverse genome hybridization, and sequenced a DNA clone, pNZE25, from a partial genomic library of the Asian rice gall midge Orseolia oryzae (Wood-Mason) (O.o.). Clone pNZE25 is highly A + T rich (67%), lacks any open reading frame and does not display homology to sequences in GenBank. Clone pNZE25 detects a 120-bp repeat in the O.o. genome, as seen from the generation...
A human genomic clone, ψGS, containing an intron-less glutamine synthetase (GS)-encoding pseudogene, was isolated by screening a human library. A sequence of 3004 bp, containing the GS coding region and both the 5 and 3 flanking sequences, was identified that exhibits all the characteristics of a processed pseudogene. The coding region shows 93% identity with the human GS cDNA (hGS) sequence...
a cDNA clone encoding the Ca 2 + -activated photoprotein, obelin (Obl), from Obelia longissima was sequenced. The nucleotide (nt) sequence contained two long overlapping open reading frames (ORFs), one of which encoded apoobelin (apoObl). The deduced amino acid (aa) sequence of apoObl revealed that this 195-aa protein has three EF-hand structures that are characteristic for Ca 2...
A new vector, pATO, was constructed for rapid cloning and analysis of eukaryotic promoters. When a recombinant pATO, carrying a promoter sequence in its multiple cloning site, was introduced into COS cells, Thy-1.2 protein was produced on the cell surface, and was easily identified by an fluorescein-conjugated anti-Thy-1.2 antibody. The intensity of the fluorescence reflected the strength of the...
A rat α-albumin mRNA encodes an 80-kDa plasma protein (approx. 20 μg/ml in adult rat serum). Its 2-kb human ortholog displays extensive ligand-binding core sequence homology, but striking divergence in the C-terminal domain.