[Isolation of mRNA from protoplasts of spore-forming bacteria and study of its translation products]. / Poluchenie mRNK iz protoplastov sporobrazuiushchikh bakterii i izuchenie produktov ee transliatsii.

Total RNA was isolated from the protoplasts of Bacillus intermedius 7P which actively produced alkaline exocellular RNAse. A fraction of polyadenylated RNA was isolated from this RNA using chromatography on poly(U) Sepharose. The total RNA and poly(A) +RNA were translated in Xenopus laevis oocytes. Bacterial exocellular RNAse was identified among the products of translation by means of electrophoretic and immunological analyses. The mRNA of RNAse secreted by B. intermedius 7P was concluded to be polyadenylated.

[An intracellular inhibitory activity of RNase secreted by Bacillus intermedius]. / Vnutrikletochnaia ingibitornaia aktivnost' RNKazy, sekretiruemoi Bacillus intermedius.

Bacillus intermedius cells producing extracellular RNAse were found to contain its inhibitor and an RNAse-inhibitor complex. Bacillus subtilis and Escherichia coli cell lysates did not inhibit the activity of homogeneous extracellular RNAse produced by B. intermedius. The inhibitor was shown to be specific for this RNAse and did not interact with other RNAses. As was demonstrated by biochemical tests and electrophoretic analysis, the inhibitor is released when the protoplasts are disintegrated, i.e. it is located in the cytoplasm. A correlation has been established between the biosynthesis of extracellular RNAse and its intracellular inhibitor.

[Isolation and study of polyadenylated RNA from Bacillus intermedius]. / Vydelenie i izuchenie poliadenilirovannoi RNK iz Bacillus intermedius.

A poly(A)+RNA fraction was isolated from the overall RNA of Bacillus intermedius using chromatography on poly(U) Sepharose and was shown to be electrophoretically heterogeneous. The presence of a polyadenylate segment was confirmed by hybridization with polyuridine. The biological activity of the poly(A)+RNA was proved by the translation in Xenopus laevis oocytes. The dynamics of poly(A)+RNA synthesis was studied in the course of B. intermedius growth and the content of poly(A)+RNA was assayed in the cells grown in different media.

[Intracellular and extracellular ribonucleases of Bacillus intermedius]. / Izuchenie vnutrikletochnykh i vnekletochnykh ribonukleaz Bacillus intermedius.

Endocellular and exocellular ribonucleases were studied in Bacillus intermedius. Two fractions of ribonucleases (Rf 0.72 and 0.96) were found to be associated with the cellular surface and seven fractions (Rf 0.1, 0.17, 0.33, 0.45, 0.72, 0.82 and 0.96) were detected in the cytoplasm. RNAase with Rf 0,096 had the highest activity and was repressed by inorganic orthophosphate. This RNase accumulated in the cell during the stationary growth phase just as the free enzyme form did in the culture medium. The immunological characteristics of these enzymes were identical as was shown by immunochemical analysis.

[Isolation and purification of alkaline ribonuclease from Bacillus intermedius]. / Metod vydeleniia i ochistki shchelochnoi ribonukleazy Bacillus intermedius.

The paper describes a method for preparing extracellular alkaline ribonuclease from Bacillus intermedius (EC 3.1.4.23). The method consists of acid treatment of the culture fluid for selective inactivation of the interfering enzymes, concentration of the enzymic protein by ammonium sulfate precipitation, dialysis against water, chromatography on DEAE-cellulose and phosphocellulose in the steady state, rechromatography on a phosphocellulose containing column, desalting and freeze-drying of the end product. Experimental samples of ribonuclease of 92% purity have been thus obtained.