Gene
Aldehyde dehydrogenases (ALDH; EC 1.2.1.3) are a family of isozymes which have been suggested to play a major role in the detoxification of aldehydes generated by alcohol metabolism and lipid peroxidation. Five non-allelic ALDH genes, encoding the ALDH1, 2, 3, 5 and 6 isozymes, have previously been identified and cloned in our laboratory. In this paper, we report the cloning and sequencing of a...
We have isolated a cDNA clone encoding the 104-kDa major vault protein (MVP) from Rattus norvegicus. The complete nucleotide sequence was determined. Comparison of the deduced amino acid (aa) sequence to that of MvpA from Dictyostelium discoideum revealed that the proteins share about 57% aa identity. Southern blot analysis indicates that the rat MVP is a single-copy gene.
Using a modification of the procedure developed by Cox et al. [Genomics 4 (1989) 397-407], we isolated and characterized 60 radiation hybrids (RH) prepared by fusing an X-ray-irradiated Chinese hamster-human chromosome 3 (Chr 3) cell line (Q314-2) with a UrdA Chinese hamster mutant cell line. The RH were screened for human DNA content by PCR amplification using primers directed to the human Alu repeat...
A nucleotide sequence of 7 kb is reported, encompassing two bleomycin-resistance (Bm R -encoding) genes and five other open reading frames (ORFs) from the Bm-producing organism Streptomyces verticillus ATCC 15003. The deduced ORFs, in sequence order, encode for (i) a protein homologous to an amino-acid dioxygenase; (ii) BlmA, the Bm R -binding protein described by Sugiyama et al....
The Rickettsia prowazekii (Rp) gyr A gene, which codes for a subunit of DNA gyrase in this obligate intracellular bacterium, has been isolated and characterized. Nucleotide sequence analysis revealed an open reading frame (ORF), initiating with a GTG start codon, of 2718 bp that could encode a protein of 905 amino acids (aa) with a calculatedM r of 101 048. The Rp gyrase subunit A (GyrA),...
The nucleotide (nt) sequence of the 5 portion of the gene encoding the Plasmodium yoelii yoelii (Pyy) 17XL merozite surface protein-1 (MSP-1) was determined by direct sequencing of both strands of the polymerase chain reaction (PCR) product. This report completes the entire coding region of the Pyy 17XL MSP-1 gene which we have found to be identical to the nt sequence of the Pyy YM MSP-1 [Lewis,...
Expression of the proU operon of Escherichia coli is directly proportional to the osmolarity of the growth medium. The basal level of proU transcription is very low, but a large increase is triggered by a sudden rise in the external osmolarity. This increased expression is maintained for as long as the osmotic stimulus persists. We have capitalized upon these regulatory features of the proU operon...
The MID2 gene whose defect (the mid2-1 mutation) results in mating-pheromone-induced death in Saccharomyces cerevisiae was cloned and its nucleotide (nt) sequence determined. The sequence showed an open reading frame (ORF) coding for a 376-amino-acid (aa) protein with an estimated M r of 39 104, and six potential TATA boxes and two pheromone-response elements in its 5 -upstream region. ...
The cloning and characterization of cDNAs encoding the cell-surface-specific integrin receptors of baboon platelets has been undertaken to provide species-specific probes. These will be used to investigate the expression and distribution of these receptors among primate species. Clones GPIIb-16 and GPIIb-3, encoding portions of the baboon glycoprotein GPIIb, were isolated from a cDNA library derived...
p-Nitrobenzyl esters serve as protecting groups on intermediates in the manufacture of clinically important oral β-lactam antibiotics; de-esterification of the intermediates is required for synthesis of the final product. A Bacillus subtilis PNB carboxy-esterase (PNBCE) catalyzes hydrolysis of several β-lactam antibiotic PNB esters to the corresponding free acid and PNB alcohol. This communication...
A series of Saccharomyces cerevisiae - Escherichia coli shuttle vectors is described in which small RNAs can be stably expressed in yeast from two different promoters for RNA polymerase III transcription. The vectors are available in either high- or low-copy-number forms with either URA3, HIS3, or TRP1 selection markers, and are based on a previously described set of plasmid vectors [Sikorski and...
A Drosophila melanogaster (Dm) embryonic cDNA library was screened for genes capable of inhibiting weel + /mik1 + protein kinase (Pk) function. We expected to identify homologs of the Schizosaccharomyces pombe gene nim1 + . This gene encodes a Pk capable of phosphorylating and so inhibiting the wee1 + Pk that in turn inhibits p34 c d c 2 . Dm cDNAs...
Fractionation of DNA from Trypanosoma brucei and T. equiperdum by centrifugation in a Cs 2 SO 4 /BAMD density gradient indicated that these genomes are compositionally compartmentalized, a conclusion confirmed by the analysis of the compositional distribution of third codon positions from T. brucei and T. cruzi. In order to investigate whether this compartmentalization is accompanied...
Plasmids containing the origin of bidirectional replication (oriβ) of the Chinese hamster dihydrofolate reductase-encoding gene (DHFR) were tested for autonomous replication in vivo and in vitro. The results show that plasmids pX24 and pneoS13, that contain a 4.8- and a 11.5-kb fragment, respectively, spanning the oriβ region, are able to replicate autonomously in human cells and in a cell-free system...
Ras proteins bound to GDP are biologically inactive while those bound to GTP are active. Ras-specific guanine nucleotide-exchange factors (GEFs) have been shown to activate Ras proteins. We used oligodeoxyribonucleotide primers with sequences similar to the cDNAs of rat and mouse cdc25 (encoding a Ras-GEF) to amplify, by the PCR, sequences with the potential to encode a 1275-amino-acid protein homologous...
The monoclonal antibody (mAb) KAA8 recognizes the peptide angiotensin II (AII). The KAA8 mAb was biopanned using two phage-displayed peptide libraries, one unconstrained, the other constrained by a disulfide bond. After several cycles of biopanning, both libraries showed enrichment for phage that bind KAA8. Phage isolated from the unconstrained library contain a consensus sequence that matches the...
The nucleotide sequence of YKE2, a yeast nuclear gene, has been determined. The deduced YKE2 protein has 114 amino acids (12 kDa), shows significant homology with the murine KE2 and contains a putative Leu-zipper motif characteristic of a group of DNA-binding proteins [Landschulz et al., Science 240 (1988) 1759-1764]. Strains in whichYKE2 has been disrupted show normal cell growth in glucose and...
We have constructed two new mini-Mu derivatives, pMRfP and pBEf, that combine the properties of known mini-Mu vectors and the advantages of the replication origin (ori f d ) of filamentous phage fd. Mini-Mu pMRfP consists of the left (850 bp) and the right (216 bp) ends of the Mu genome, ori f d , packaging signal of fd, and the gene conferring resistance to chloramphenicol...
The dps genes of Streptomyces peucetius, encoding daunorubicin (DNR)-doxorubicin (DXR) polyketide synthase (PKS), are largely within an 8.7-kb region of DNA that has been characterized by Southern analysis, and gene sequencing, mutagenesis and expression experiments. This region contains nine ORFs, many of whose predicted products are homologous to known PKS enzymes. Surprisingly, the gene encoding...
The macromolecular synthesis (MMS) operon consists of three genes: rpsU, which encodes the S21 ribosomal protein (in Bacillus subtilis (Bs), rpsU is replaced by orfP23 which encodes a protein of unknown function), dnaG, encoding the DNA primase involved in the initiation of chromosome replication, and rpoD, which encodes the principal sigma subunit of RNA polymerase. The operon was cloned in three...