Advanced practice nursing: the winds of change.
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Humans↗
Biomedical subjects
G L Ray
Publications and source records attributed to G L Ray.
Advanced practice nursing: playing a vital role.
To clarify the education preparation of advanced practice nurses, a survey of all 50 states was performed. Results show a high degree of variability from state to state in tracking advanced practice nurses. Standardizing the educational preparation for the nurse practitioner, clinical nurse specialist, certified nurse midwife and the certified registered nurse anesthetist will clarify advanced practice nursing.
Clinical Competence↗
Nucleotide sequences that define promoters that are used by Bacillus subtilis sigma-29 RNA polymerase.
There are at least five different forms of RNA polymerase holoenzyme in Bacillus subtilis. These enzymes differ in their sigma subunit and their specificity for promoter utilization. One form of RNA polymerase (E sigma 29) that contains a 29,000 Mr sigma appears in B. subtilis about two hours after the initiation of endospore formation. The determination of the nucleotide sequences that govern utilization of promoters by E sigma 29 has been limited by the small number of cloned promoters that are recognized by E sigma 29. We have determined the nucleotide sequence of a recently isolated promoter (G4) that is used exclusively by E sigma 29 both in vitro and in vivo. The start-point of transcription was identified by S1 nuclease mapping and dinucleotide priming experiments and the probable promoter element was sequenced. We compared the sequence with that of six promoters that are used to varying degrees in vitro by E sigma 29 and found these sequences to be highly conserved at the -10 and near the -35 regions of these promoters. Single base substitutions were generated at positions -12, -15 and -36 of the G4 promoter and assayed for their influence on utilization by E sigma 29 in in-vitro competition experiments. The effects of these mutations in G4 on its use by E sigma 29 support a model in which E sigma 29 utilizes its cognate promoters by interacting with unique nucleotide sequences at the -10 region and near the -35 region of these promoters.
Bacillus subtilis↗
Isolation of Bacillus subtilis genes transcribed in vitro and in vivo by a major sporulation-induced, DNA-dependent RNA polymerase.
As a means of determining the function of sigma 29, a sporulation-essential sigma factor, we have isolated and begun to characterize genes that require sigma 29 for their expression. RNA transcribed in vitro from total Bacillus subtilis DNA by using sigma 29-containing RNA polymerase (E-sigma-29) was hybridized to a bank of B. subtilis DNA fragments that had been cloned into bacteriophage lambda. Approximately 0.25% of the cloned B. subtilis DNA fragments displayed detectable hybridization with our RNA probe. Five DNA fragments that had strong in vitro template activity for E-sigma-29 were selected for further study. The DNA fragments which contained in vitro sigma 29 promoter activity encoded RNAs that were synthesized by B. subtilis during sporulation. Mutant B. subtilis that failed to synthesize sigma 29 (spoIIA, spoIIE) made less RNA that could hybridize to these cloned DNAs than did a mutant (spoIIC) which did synthesize sigma 29 but was blocked at a similar stage in development. A detailed analysis of several of the cloned DNAs demonstrated that they encoded RNAs that were transcribed from approximately the same start site in vivo that E-sigma-29 initiated transcription in vitro. These particular transcripts were present only during the period of sigma-29 abundance (2 to 4 h after the onset of sporulation) in sporulating cells which carried a wild-type allele of the sigma-29 structural gene (spoIIG). We conclude that the isolation procedure used in this study identified genes that are transcribed by E-sigma 29, not only in vitro but also in vivo. Preliminary characterization of the cloned genes indicate that they encoded multiple overlapping RNAs which were each synthesized at unique times during growth or sporulation. This result implies that sigma 29 does not activate a unique population of genes with a novel function in sporulation but rather that it has a temporal role in spore gene control, transcribing those genes required to be active during its period of abundance regardless of their specific function.
Bacillus subtilis↗
Isolation of Bacillus subtilis mutants altered in expression of a gene transcribed in vitro by a minor form of RNA polymerase (E-sigma 37).
To develop a technique for identifying Bacillus subtilis genes whose products affect transcription from promoters recognized by sigma 37-containing RNA polymerase (E-sigma 37), we cloned the promoter region of a gene (ctc) that is actively transcribed in vitro by E-sigma 37 into a plasmid (pPL603B) so that a transcriptional fusion was created between ctc and a plasmid-borne chloramphenicol acetyltransferase (CAT) gene. CAT levels in B. subtilis carrying the ctc/CAT fusion plasmid varied in a manner that was consistent with the known pattern of ctc RNA synthesis. Mutagenesis of cells harboring the ctc/CAT plasmid led to the isolation of bacterial clones which displayed altered chloramphenicol resistance. Analysis of the mutants demonstrated that CAT activity was substantially changed in the mutant cells. Several of the B. subtilis mutants, both CAT overproducers and underproducers, also had acquired a sporulation-deficient phenotype. The mutations responsible for altered CAT expression were not carried on the plasmid. Analysis of RNA synthesized by mutant cells indicates that at least a portion of the mutants may be altered in the level of transcription from the ctc promoter and, hence, are likely to define B. subtilis genes which influence this process.
Acetyltransferases↗