Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:Q9UIJ5 (Rec)
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We have demonstrated in this study that CaCl2-MnCl2 treatment is effective in the preparation of competent cells of Erwinia chrysanthemi for transformation. Plasmids pMB9 and pBR322, which are of Escherichia coli origins, were transformed into E. chrysanthemi at frequencies of 1.5 X 10(-7) and 4.5 X 10(-7) per recipient, respectively. The frequencies were 60- to 80-fold lower than those in the well established rec- E. coli; however, the procedure is practically useful in transformation experiments. The plasmids were maintained rather stable in the cells if antibiotics were included in the media. When transformed by pECl and pEC6, which were chimeric plasmids consisting of pBR322 and an E. chrysanthemi chromosomal DNA fragment of 2.4 and 3.5 Mdal, respectively, the transformation frequencies of E. chrysanthemi were at the same range of that by pBR322. Neither colony morphology nor the polypectate degradation ability was changed after transformation by the plasmids. In conclusion, we have established a system of molecular cloning in E. chrysanthemi SR120A exploiting pBR322 as a vector.
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PMID:Transformation of Erwinia chrysanthemi by Escherichia coli plasmids DNA. 389 40

Small-granule endocrine cells differentiate in airway epithelium of intact and cultured fetal rat lungs. We noted that the cells store calcitonin gene-related peptide (CGRP) in vitro as well as in vivo and used the ionophore A23187 to test the effects of calcium on peptide secretion in this system. Lungs of 14-day and 15-day fetal rats, organ cultured for 6-9 days, were divided into groups of 5 explants each and incubated for 15 min at 37 degrees C in the basic medium containing 0 mM, 1 mM, or 10 mM CaCl2, with or without 8 microM A23187, or 10 mM EGTA. Intracellular CGRP in these explants was quantified by supraoptimal dilution peroxidase immunocytochemistry (Springall et al.: J. Pathol. 155:259-267, 1988): counts were made of endocrine cells stained with a 1/60,000 dilution of anti-CGRP and repeated on the same sections after restaining with antibody diluted at 1/1,000. Results, analyzed by Chi-square test, were expressed as % cells stained with antibody at 1/60,000 vs. those stained at 1/1,000. Immunoreactivity for CGRP was significantly reduced by A23187 in the presence of high extracellular Ca2+ (10 mM), the inference being that these cells secrete peptide hormones in response to Ca2+ influx across the plasma membrane. The organ cultures evidently can be used to assess certain physiological responses of lung endocrine cells in an accessible, relatively organotypical setting.
Anat Rec 1993 May
PMID:Calcium and ionophore A23187 lower calcitonin gene-related peptide-like immunoreactivity in endocrine cells of organ cultured fetal rat lungs. 850 10