Direct selection for curing and deletion of Rhizobium plasmids using transposons carrying the Bacillus subtilis sacB gene

We have constructed derivatives of the transposon Tn5 carrying the mob site (oriT) of plasmid RP4, and an nptI-sacB-sacR cassette [Ried and Collmer, Gene 57 (1987) 239-246]. The mob site, in conjunction with the antibiotic-resistance markers carried on the transposons, allows identification of transposon inserts in cryptic plasmids by mobilisation to other strains. The sacB-sacR genes allow direct selection for the loss or curing of plasmids, because only strains which no longer contain an active sacB gene are able to grow on media containing sucrose. We have tested these transposons in four strains of Rhizobium leguminosarum and two strains of Rhizobium meliloti, and have been able to demonstrate curing of several large cryptic plasmids, and generation of large deletions in many other plasmids. This method has enabled us to show that the R. leguminosarum plasmids pRL12JI and pR1eVF39f carry auxotrophic markers, and that the plasmid pR1eVF39c carries genes which affect colony morphology.     

The control of oscillatory movements of the forearm

The patterns of EMG activity in the biceps and triceps muscles were recorded during horizontal oscillatory movements of the forearm. Subjects showed increased frequency of oscillation as they voluntarily reduced movement amplitude. EMG burst duration was significantly correlated with wavelength of oscillation in every case. In almost half the cases burst intensity was also positively correlated with wavelength. Subjects seemed to be using one or both these methods to control amplitude. A model was developed in three stages which satisfactorily accounted for the data.     

Three types of vitronectin in human blood

Vitronectin is a cell-adhesive glycoprotein in serum and plasma, also termed serum spreading factor and complement S-protein. It consists of a mixture of a polypeptide of molecular weight 75 kilodalton (kDa) and its nicked product of 65 kDa plus 10 kDa. By a quantitative immunoblotting assay, human blood samples could be classified into three distinct vitronectin types; type I (58% of the population) was 75 kDa rich and 65 kDa poor, type II (35% of the population) contained approximately equal amounts of 75 kDa and 65 kDa, and type III (5% of the population) was 75 kDa poor and 65 kDa rich. The vitronectin type did not correlate with age, sex, or ABO blood type.     

A radio-gas chromatographic method for determining the specific radioactivity of glycolic acid in -14C-labeled leaf tissue.

A method for the extraction and quantitative determination of both the mass and radioactivity of glycolic acid from -14C-labeled leaf tissue is described. The recoveries of both mass and radioactivity from standard [1-14C]glycolic acid solutions averaged 98 percent, and recovery of radioactivity added to plant samples as [1-14C]glycolic acid was over 90 percent after the complete procedure. The method was reliable with total samples containing as little as 130 nmol of glycolic acid. The mass of glycolic acid recovered from sunflower leaf tissue was proportional to the amount of tissue extracted. In experiments with different plant material, the amount of glycolic acid varied between 530 and 1120 nmol/dm-2 of leaf tissue. The specific radioactivity of the glycolic acid in sunflower leaf tissue during photosynthesis in -14CO(2) was never more than 20 percent of the specific radioactivity of the -14CO(2) supplied.     

Intermittent illumination increases biophotolytic hydrogen yield by Anabaena cylindrica.

Intermittent illumination increased H2 and C2H4 yields per unit of light from growing cells and from nitrogren-starved cells by 1.7- and 1.35-fold, respectively, as compared with continuous illumination.     

The release of alanine by rat diaphragm muscle in vitro.

Z discs were isolated from Lethocerus flight muscle by removing the contractile proteins from myofibrils with a solution of high ionic strength. The protein composition of the Z discs was analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; the major proteins were alpha-actinin, actin and tropomyosin. Z lines were selectively removed from intact myofibrils by digestion with crude lipase and chymotrypsin, but not by purified lipase.