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At least 451 records · Page 25Linked to original sources
PubMed1971

Morphogenetic aspects of murein structure and biosynthesis.

The shape of Escherichia coli is fixed by the form of the sacculus. This sacculus is a macromolecule made up from the polymer murein. In an investigation of the possible factors determining the shape of the sacculus, we attempted to resolve between two fundamental alternatives. (i) Is the shape of the sacculus automatically fixed by its chemical composition? or (ii) does a special morphogenetic system exist which determines the shape of the sacculus? An analysis of sacculi from cells grown in poor and rich media and harvested at different stages of growth was made. Significant variations in the composition of murein were found, whereas the general shape of the cells remained unchanged. This finding stands opposed to the assumption of a strict correlation between chemistry and shape of the sacculus. The second alternative was investigated by attempting to change artificially the shape of the sacculus by modifying the form of the hypothetical morphogenetic system. Rod-shaped cells were converted into spherical spheroplasts which were subsequently allowed to reform a new spherical sacculus. In chemical composition this spherical sacculus was found to be indistinguishable from the rod-shaped sacculus. This finding is taken as evidence for the existence of a distinct morphogenetic apparatus in the cell wall whose form is reflected by the shape of the sacculus.

Bacterial Proteins↗
PubMed1973

Resistance of Escherichia coli to penicillins: identification of the structural gene for the chromosomal penicillinase.

A screening procedure was used to isolate a number of mutants of Escherichia coli K-12 with low penicillinase activity. By co-transduction with purA, three of the mutants were found to map near 82 min. Penicillinase was purified from one mutant and from a transductant with a temperature-sensitive enzyme. Comparison with wild-type penicillinase revealed similarities in the Ouchterlony immunodiffusion test but differences in the catalytic properties. It is concluded that the mutations have occurred in the structural gene of the chromosomal penicillinase (designated ampC). Purified enzyme and a temperature-sensitive mutant were used to investigate whether the penicillinase has a physiological function related to biosynthesis or breakdown of murein. No positive evidence for any such function was obtained.

Chromosome Mapping↗
PubMed1975

Genetics of biotin biosynthesis in Bacillus subtilis.

Biotin auxotrophs of Bacillus subtilis were isolated and classified into three groups according to growth requirements, cross-feeding pattern, and biotin precursors excreted into culture supernatant fluids. Mutant genes were mapped by transduction using phage PBS1. All presently identified bio genes were linked to aroG with an order of bio-aroG-argA-leu-1. No linked markers were found to the left of the bio loci.

Arginine↗
PubMed1969

Immunochemistry of the cell walls of Listeria monocytogenes.

The antigenic specificity of Listeria monocytogenes types I, II, III, IVa, and IVb was studied by immunochemical techniques. Immunologically active carbohydrates of the various types were extracted from cell walls and were chemically analyzed. Types I and II contained predominantly glucosamine and rhamnose; type III, galactose, rhamnose, and glucosamine; and types IVa and IVb, glucose and galactose. Quantitative precipitin inhibition tests with purified monosaccharides indicated that the major antigenic determinant of types I and II is rhamnose. Precipitin reactions could not be detected with type III carbohydrate and homologous or heterologous antisera. The major determinants of types IVa and IVb were found to be galactose and glucose, respectively. As much as 87% inhibition of the quantitative precipitin test for types I and II was obtained with rhamnose, 72% for type IVa with galactose, and 72% for type IVb with glucose. The immunochemical basis for the antigenic specificity of L. monocytogenes types I, II, IVa, and IVb was further confirmed by using agar gel diffusion. Cross-reactions among the various type-specific carbohydrates and heterologous antisera were also studied. Type II carbohydrate was found to contain galactose and react with type IVa antisera. This reaction could be blocked by galactose. Type I carbohydrate did not contain galactose nor did it react with antiserum prepared from type IVa cells. Therefore, the somatic antigens of type I and type II L. monocytogenes, previously thought to be identical, appeared to differ. The dominant immuno-specific group in the cross-reaction between type IVb carbohydrate and type IVa antisera was found to be galactose. Type IVa absorbed antisera did not produce a significant cross-reaction with type IVb carbohydrate. The results obtained from this investigation indicate a lesser degree of antigenic relationship between type IVa and type IVb L. monocytogenes than was previously believed to exist.

Alanine↗
PubMed1974

Evidence for the presence of nontranslated T7 late mRNA in infected F'(PIF+) episome-containing cells.

The inability of T7 to develop in cells of Escherichia coli containing F(+) or substituted F' episomes is a result of the failure to synthesize late proteins; no in vivo translation of mRNA species synthesized by the T7 RNA polymerase occurs. Further experiments have been performed to measure the amount of late mRNA in T7-infected F'(PIF(+)) cells. (We have designated the property of phage inhibition of F factors as PIF; the wild-type episome is therefore F'[PIF(+)].) T7 late proteins were synthesized in vitro by using a system programed with RNA extracted from T7-infected F(-) and F'(PIF(+)) cells. The T7 lysozyme, product of gene 3.5, and the gene 10 head protein were assayed. The following results were obtained: (i) mRNA capable of supporting in vitro synthesis of lysozyme and the gene 10 head protein is present in T7-infected F'(PIF(+)) cells; (ii) lysozyme mRNA extracted from T7-infected F'(PIF(+)) cells is present at 70 to 75% of the level found in T7-infected F(-) cells; (iii) gene 10 mRNA is present at 35 to 78% of the level found in T7-infected F(-) cells. No in vivo synthesis of either lysozyme or gene 10 protein can be detected in T7-infected F'(PIF(+)) cells although normal synthesis of these proteins occurs in F(-) cells. These findings confirm that the block in T7 development in F'(PIF(+)) cells results from the failure to translate late classes of T7 RNA.

Cell-Free System↗
PubMed1990

[Studies on the induction of the humoral immune responses to Bacteroides gingivalis fimbrial antigen in mice].

Serum and salivary antibody responses to Bacteroides gingivalis fimbriae administered either orally or subcutaneously (s.c.) with or without an adjuvant in various strains of mice were examined in this study. Following results were obtained. 1) Oral administration of B. gingivalis fimbriae with GM-53 as an adjuvant in liposomes, but not in Tris-HCl buffer, definitely enhanced the fimbriae-specific IgG responses, mainly IgG1 followed by IgG2b, IgG2a and IgG3 in serum and IgA response in saliva of BALB/c mice. On the other hand, s.c. injection of fimbriae with GM-53 or MDP-Lys (L18) also raised the fimbriae-specific IgG followed by IgA and IgM responses in serum, and both IgA and IgG responses in saliva of BALB/c mice. Oral immunization was less effective than s.c. injection in terms of the production of serum antibody in the mice. However, the level of salivary antibody of mice injected s.c. was similar to that of mice immunized orally. 2) High anti-fimbriae antibodies in serum were maintained in BALB/c mice immunized orally with fimbriae and GM-53 in liposomes for approximately 7 months after the primary immunizations. Oral administration also induced and held the fimbriae-specific IgA response in saliva for at least 6 months after the primary immunizations. The levels of fimbriae-specific IgA in saliva after the second boosters on days 123 and 124 were higher than those after the primary ones on days 27 and 28. 3) Among various strains of mice immunized orally with fimbriae and GM-53 in liposomes, BALB/c and DBA/2 mice (H-2d) significantly produced high levels of both serum IgG and salivary IgA antibodies specific for fimbriae. Furthermore, B10.D2 mice (H-2d) were responders followed by B10.BR (H-2k), while C57BL/10 mice (B10, H-2b) were low responders to the fimbriae. These results show that the combined use of fimbriae together with an adjuvant results in a sharply increased IgA antibody response in saliva and a predominantly stimulated IgG antibody in serum, and it was suggested that these responses are restricted by H-2 haplotype.

Acetylmuramyl-Alanyl-Isoglutamine↗