Intraspecific and intergeneric mobilization of non-conjugative resistance plasmids by a 24.5 megadalton conjugative plasmid of Neisseria gonorrhoeae

pLE2451, a 24.5 megadalton conjugative plasmid from Neisseria gonorrhoeae, was capable of efficiently mobilizing gonococcal beta-lactamase plasmids between gonococci and from gonococci to Haemophilus influenzae and restriction-deficient Escherichia coli. Donor strains of N. gonorrhoeae carrying pLE2451 were also found to be capable of mobilizing a variety of non-conjugative plasmids originally derived from enteric bacteria or Haemophilus species when such plasmids were resident in E. coli. Nevertheless, pLE2451 was not detected physically in E. coli or H. influenzae transconjugants. This suggests that the plasmid is unstable in these hosts but survives transiently to provide transfer functions for mobilization. The proficiency of pLE2451 in promoting intraspecific and intergeneric mobilization was not paralleled by pUB701, pRI234 and pFR16017, a series of conjugative plasmids derived originally from Haemophilus species. These plasmids were incapable of mobilizing even Haemophilus beta-lactamase plasmids, such as RSF0885, between Haemophilus species.     

Pathway of plasmid transformation in Pneumococcus: open circular and linear molecules are active.

We have extended the analysis of plasmid transformation in Streptococcus pneumoniae by finding that monomeric and dimeric open circular and linear forms of pMV158 were active in transformation. Their efficiencies were at least 35-fold lower than those of the corresponding closed circular forms. The evidence came largely from analysis of S1 nuclease-digested plasmid deoxyribonucleic acid by combinations of dye-buoyancy, gel electrophoresis, and sedimentation velocity methods. As with closed circular forms, monomer open circular forms gave second-order kinetics and dimer forms gave first-order kinetics. Unique linear products of digestion by either of two restriction enzymes were inactive, but a mixture of the two digests was active, as was the mixture of linear monomer deoxyribonucleic acids produced by S1 nuclease. Absolute efficiencies of transformation were low even for closed circular donors. All of the results, including the low efficiencies, were consistent with the interpretation that plasmid replicons were assembled in the recipient cell by pairing of fragments of single strands that had entered the cell separately from duplex donors that had been cut on the cell surface.     

Preparation of pancreatic mRNA: cell-free translation of an insulin-immunoreactive polypeptide.

Total nucleic acid extraction and selection of poly A-containing molecules yield preparative quantities of undegraded mRNA from adult and fetal pancreas. Using a stringent immunoassay, this mRNA is found to direct the synthesis of an immunoreactive insulin polypeptide in the wheat germ translation system. On sodium dodecyl sulfate polyacrylamide gels, this polypeptide (12,000-13,000 daltons) is larger than proinsulin (9,000 daltons).     

The effect of 3-mercaptopicolinic acid on phosphoenolpyruvate carboxykinase (GTP) in the rat and guinea pig.

In vitro, 3-mercaptopicolinic acid inhibited phosphoenolpyruvate carboxykinase activity in supernatant fractions of liver, kidney cortex, and adipose tissue obtained from fasted rats. 3-Mercaptopicolinic acid also inhibited enzymatic activity in the mitochondrial and supernatant fractions of liver obtained from fasted guinea pigs. In the fasted rat, the oral administration of 3-mercaptopicolinic acid increased liver carboxykinase activity even though the blood glucose concentrations decreased. Kidney cortex carboxykinase decreased while adipose tissue enzyme was unchanged. In the fasted guinea pig, the oral administration of 3-mercaptopicolinic acid lowered blood glucose concentrations but had no effect on liver mitochondrial or supernatant carboxykinase activity. The elevation in rat liver enzymatic activity appears to be due to protein synthesis, since the concurrent administration of cycloheximide prevents the increase in enzyme activity. 3-Mercaptopicolinic acid appears to be noncompetitive with respect to Mn²⁺.     

Albumin synthesis and catabolism following partial hepatectomy in the rat. The effects of amino acids and adrenocortical steroids on albumin synthesis after partial hepatectomy.

Plasma albumin levels were measured in partially hepatectomized, sham operated and control rats. The levels fell in both the partially hepatectomized and sham operated groups; while the latter group returned to normal within a few days, the low plasma albumin in the partially hepatectomized animals was sustained. Albumin synthesis rates in the isolated perfused rat liver were measured in the three groups of animals at varying intervals after partial hepatectomy. There was a significant depression of albumin synthesis rate in terms of both liver and whole animal weights when compared to the sham operated and control animals. This depression was almost completely reversed by the addition of arginine, asparagine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan and valine added together to 10 times their normal plasma concentrations. The addition of hydrocortisone had no effect on the albumin synthesis rate after partial hepatectomy. Studies in vivo in the three groups of animals (partially hepatectomized, sham operated and control animals) revealed a fall in the albumin catabolic rate after partial hepatectomy coinciding with the fall in the albumin synthesis rate. An hypothesis whereby the amino acids may have their stimulatory effect is proposed.     

Inductive activity and enduring cellular constitution of a supernumerary apical ectodermal ridge grafted to the limb bud of the chick embryo.

Apical ectodermal ridges (AERs) isolated from 3- to 4-day chick and quail embryos were prepared by means of trypsinization and microdissection and then were grafted to the dorsal or ventral side of a host chick wing bud. They induced supernumerary limb outgrowths from the host bud showing, respectively, a bidorsal or biventral organization, as determined by the patterns of feather germs. The grafted ridge cells persisted, as revealed by histological sections of supernumerary chick limb parts growing under the influence of quail AERs, whose cells are readily distinguished after application of the Feulgen reagent.These results show that the AER induces limb outgrowth regardless of whether it is associated with dorsal or ventral limb ectoderm and that its continued existence is not dependent on contributions of ectodermal cells from the opposed ectodermal faces of the limb bud. The AER is pictured as maintaining the subjacent mesoderm in a condition of developmental plasticity without specifying its differentiation with respect to the proximodistal axis. It remains uncertain whether the positional values of cells that develop under the influence of the AER arise within these cells themselves or appear in response to influences from proximal sources.     

Extrachromosomal deoxyribonucleic acid in wild-type and photosynthetically incompetent strains of Rhodopseudomonas spheroides.

Three covalently closed circular species of extrachromosomal deoxyribonucleic acid have been identified by electron microscopic analysis in strains of Rhodopseudomonas spheroides. The weights of these plasmids, as determined from contour length, are about 75 X 10(6), 66 X 10(6), and 28 X 10(6) daltons for both aerobically grown and photosynthetically grown R. spheroides strain 2.4.1 (NRS) and for the photosynthetically incompetent strain V-2 (obtained by N-methyl-N-nitro-N'nitrosoguanidine mutagenesis) and 74 X 10(6), 66 X 10(6) and 34 X 10(6) daltons for a second photosynthetically incompetent strain, SLS I (obtained by incubating strain 2.4.1 [NRS] in medium containing sodium lauryl sulfate). Buoyant densities uere found to be 1.717 g/cm3 (58% guanine plus cytosine) for the plasmids of 66 X 10(6), 28 X 10(6), and 34 X 10(6) daltons in weight and 1.724 g/cm3 (65% guanine plus cytosine) for those weighing about 75 X 10(6) daltons. Possible functions of these plasmids are discussed.