[Creation of the library of Bordetella pertussis genome clones and detection of clones containing genes for the synthesis of antigenic determinants of the whooping cough microbe]. / Sozdanie klonoteki genoma Bordetella pertussis i obnaruzhenie v nei klonov, soderzhashchikh geny sinteza antigennykh determinant kokliushnogo mikroba.

Escherichia coli clones containing hybrid phasmids with the inserts of B. pertussis DNA were obtained with the use of a phasmid vector. The total amount of the clones thus obtained was 97,000, which considerably exceeded the volume of the clone library necessary for the detection of individual genes with probability approximating 1. The hybrid plasmids were shown to contain 6-19 kilobases. The screening of the clone library was carried out by means of the enzyme immunoassay (EIA). The assay was aimed at detecting clones containing the genes of the subunits of B. pertussis lymphocytosis-stimulating factor (LSF). The EIA techniques used in this investigation were based on the capacity of LSF for binding with fetuin. Six clones giving positive response were detected. These data suggest the presence and expression of the genes controlling the synthesis of the antigenic determinants of LSF in E. coli cells.

[Isolation and purification of restriction endonuclease PmiI from Proteus mirabilis 1667]. / Vydelenie i ochistka restriktsionnoi éndonukleasy PmiI iz shtamma Proteus mirabilis 1667.

A new restriction endonuclease Pmi I was detected in Proteus mirabilis 1667. The enzyme hydrolyzes DNA of the phage lambda into 10 electrophoretically separating fragments with molecular weights of 1.3-7.9 mD. With the use of two-stage chromatography on blue sepharose and phosphocellulose it is possible to obtain restriction endonuclease Pmi I free of the admixtures of ballast proteins, nonspecific nucleases and phosphatases.

[Production of immobilized SalI and PvuII restrictase preparations]. / Poluchenie preparatov immobilizovannykh restriktaz SalI i PvuII.

Conditions for the immobilization of specific endonucleases Sal I and Pvu II on BrCN-activated Sepharose 4B have been selected. Some physico-chemical properties of the preparations of immobilized restrictases Sal I and Pvu II have been characterized. The specific and general activity values of the preparations thus obtained have been established. The immobilized enzymes have been used for the multiple restriction of the DNA of phage lambda and the DNA of Neisseria meningitidis.

[Use of different media for growing the producer of the restriction enzyme XBA I]. / Ispol'zovanie razlichnykh sred dlia vyrashchivaniia produtsenta restriktiruiushchego fermenta XBA I.

The possibility of using culture media prepared from local ingredients and intended for growing the producers of restricting enzyme Xba 1 has been demonstrated. The yield of restricting enzyme Xba 1 per g of crude biomass, obtained with the use of peptone-yeast medium prepared from ingredients supplied by Difco Laboratories (USA), has proved to be 4 times greater than that obtained with the use of peptone-yeast medium prepared from local ingredients. At the same time the use of casein-saline medium ensures the yield of the enzyme, similar to that obtained with the use of peptone-yeast medium prepared from ingredients supplied by Difco Laboratories, but with a greater content of nonspecific nucleases.