Construction of high, intermediate and low-copy-number promoter-probe plasmids for Bacillus subtilis.

We constructed promoter-probe plasmids for Bacillus subtilis based on the promoterless gene for penicillinase (penP) of Bacillus licheniformis. A Bacillus stearothermophilus DNA fragment, which contained translational stop codons for all three reading frames and transcription terminators of the alpha-amylase gene, was inserted upstream from the Shine-Dalgarno sequence for the penP gene. A low-copy-number plasmid (about 10 copies per chromosome), pPF101, carried tetracycline-resistance gene, whereas another high-copy-number plasmid (about 55 copies per chromosome), pPF201, conferred resistance to kanamycin.

Reducing terminals of urinary chondroitin sulfate.

Human urinary chondroitin sulfate isolated from the cetylpyridinium chloride-complex of the non-dialyzable fraction of the pooled urine was subjected to ethanol fractionation, successive enzymic digestion with neuraminidase and mucopolysaccharidases, and anion exchange chromatography. The gas liquid chromatographic analyses of the acetyl and butaneboronic acid ester derivatives of the reduced terminal sugar units after treatment with sodium borohydride plus hydrolysis revealed that 42% of the urinary chondroitin sulfate was bound to peptide through xylose. The reducing terminal sugar units of the peptide-free form consisted of 34.

Demonstration of chondroitin sulfates degrading endo-beta-glucuronidase activity in rabbit liver.

Reduced chondroitin sulfate was incubated with rabbit liver extracts followed by reduction once more with sodium [3H]borohydride, and then passed through a Sephadex G-100 column. Chondroitin sulfate obtained from the incubation medium at pH 4 was only slightly depolymerized and was highly radioactive. Paper chromatographic analyses showed that glucuronic acid residues became exposed at the reducing terminal of chondroitin sulfate after incubation with the liver extracts.

The application of the Milner-Avigad method for the quantitative determination of endouronidase activities.

The Milner-Avigad method (Milner, Y. and Avigad, G. (1967) Carbohydr.

Erythropoietic protoporphyria--report of a family and clinical review.

Three cases of erythropoietic protoporphyria are reviewed. The three patients constitute members of one family. The protoporphyrin content of the red blood cells was high, but porphyrins and their precursors in the urine and faeces were not excessive.