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We identified and cloned the rabbit SP-B gene, encoding the pulmonary surfactant-associated protein, and sequenced its upstream region from -2635 to +428, including a much larger fragment of the upstream region than has previously been reported for an SP-B for any species. Rabbit SP-B showed substantial homology to its human counterpart in the coding and noncoding regions immediately upstream from...
The control elements of the tetracycline-resistance operon encoded in Tn10 of Escherichia coli have previously been utilized by Gossen and Bujard [Proc. Natl. Acad. Sci. 89 (1992) 5547-5551] to establish an efficient transcription regulatory system in mammalian cells. We describe here a modification of this vector system that encodes all required elements, including a neomycin-resistance gene, on...
In eubacteria, the clustering of DnaA boxes around the dnaN (β subunit of DNA polymerase III) and dnaA genes usually defines the chromosome replication origin (oriC). In this study, the dnaN locus from the cyanobacteriumSynechococcus sp. strain PCC 7942 was sequenced. The gene order in this region is cbbZp-dnaN-orf288-purL-purF which contrasts with other eubacteria. A cluster of eleven DnaA boxes...
Plasmid pBRINT is an efficient vector for chromosomal integration of cloned DNA into the lacZ gene of Escherichia coli [Balbas et al., Gene 136(1993) 211-213]. A family of related plasmids containing different antibiotic-resistance markers (Cm R or Gm R or Km R ) and a larger multiple cloning site (MCS) has been constructed. This set of plasmids, whose integration efficiencies...
Many different organisms, ranging from bacteria and fungi to fireflies and fish, are endowed with the ability to emit light, but the bioluminescent systems are not evolutionarily conserved: genes coding for the luciferase proteins (Lase) are not homologous, and the luciferins are also different, falling into many unrelated chemical classes. Biochemically, all known Lase are oxygenases that utilize...
Diseases caused by Mycobacterium tuberculosis, M. leprae and M. avium, cause significant morbidity and mortality worldwide. Effective treatments require that the organisms be speciated and that drug susceptibilities for the causative organisms be characterized. Reporter phage technology has been developed as a rapid and convenient method for identifying mycobacterial species and evaluating drug...
We have developed lac repressor-regulated retroviral expression vectors that are induced by β-galactosides upon stable transduction in mammalian cells. These vectors, derived from a Moloney-virus-based vector LNCX, contain an internal Rous sarcoma virus (RSV) or cytomegalovirus (CMV) promoter, coupled with 2 to 4 lac operator sequences and placed in anti orientation relative to the retroviral long...
A set of vectors for Gram-positive bacteria was constructed with a new feature which enables the switching down of their copy number per cell. These vectors carry the replication region of pAMβ1, containing a gene essential for replication, repE, and its regulator, copF. The latter gene was inactivated by inserting a linker into its unique KpnI site. Since copF downregulates the expression of repE,...
The baculovirus expression system using the Autographa californica nuclear polyhedrosis virus (AcNPV) has been extensively utilized for high-level expression of cloned foreign genes, driven by the strong viral promoters of polyhedrin (polh) and p10 encoding genes. A parallel system using Bombyx mori nuclear polyhedrosis virus (BmNPV) is much less exploited because the choice and variety of BmNPV-based...
In humans, at least three types of inositol (1,4,5)-trisphosphate receptor (IP 3 R) are present. The gene encoding type 1 IP 3 R (IP 3 R-I) is expressed in all cell types, although expression predominates in Purkinje cells. To study the regulation of the human IP 3 R-I gene, we isolated and characterized a 2.1-kb 5 flanking region. In transient expression assays using...
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