The immediate vicinity of mouse metallothionein-I gene contains two sites conferring glucocorticoid inducibility to the heterologous promoter.

Glucocorticoid responsive elements (GREs) located -252 to -209 by upstream and +1011 to +1054 bp downstream of the transcription initiation site of the mouse metallothionein-I (mMT-I) gene were identified in transient experiments. However, the promoter region of the mMT-I gene (-330 to +70 bp) was found to provide low, if any, glucocorticoid induction of the linked CAT gene, while showing strong cadmium regulation, comparable with the in vivo level.

Molecular cloning and sequencing of omul prolactin cDNA.

We constructed a complementary DNA (cDNA) library from Baikal omul (Coregonus autumnalis migratorius Georgi) pituitary glands. Synthetic oligonucleotide probes corresponding to chum salmon prolactin (PRL) cDNA were used to select the recombinant plasmids carrying the omul PRL gene. The larger insert was sequenced and found to encode a polypeptide of 210 amino acid residues, including a putative signal sequence of 23 amino acids.

[An experimental infection of the seal Phoca sibirica with the morbilli virus].

A Baikal seal (Phoca sibirica) was experimentally infected with Baikal seal morbillivirus (BSMV) isolated from Baikal seals during an epizootic in 1987-1988. The seal was infected with BSMV with an infectious titer of 10(7.0) TCD50/ml, and daily observations of the animal clinical condition were made.

[Antibodies to morbilli virus of the Baikal seal in its natural host].

Morbillivirus of Baikal seal (BSM) was isolated from organs of a dead animal during 1987-1988 epizootic of Baikal seal (Phoca sibirica). A method of cellular enzyme immunoassay for testing for virus-specific antibodies was developed using BSM. The method was used for antibody detection in sera of 115 apparently normal seals collected in the spring of 1989.

[Identification of the glucocorticoid receptor binding site at the 5'-flanking region of mouse metallothionein I gene: the effect of base substitutions on binding efficiency].

Interaction of highly purified glucocorticoid receptor complex (GIRC) with synthetic DNA-fragment of mouse metallotionein 1 gene promoter from -209 to -252 b.p. (MTwt) was investigated.

Comparison of two morbilliviruses isolated from seals during outbreaks of distemper in north west Europe and Siberia.

Recently morbilliviruses were isolated from harbour seals (Phoca vitulina) in North West Europe (phocid distemper virus-1: PDV-1) and from Baikal seals (Phoca sibirica) in Siberia (phocid distemper virus-2: PDV-2) during outbreaks of severe disease which resembled distemper in dogs. PDV-1 and PDV-2 were passaged in SPF dogs, in which they caused distemper-like disease symptoms, and were subsequently passaged in Vero cells in which they caused cytopathic changes. PDV-1, PDV-2, and canine distemper virus (CDV) were compared with respect to their biological, morphological, physical, protein chemical, and antigenic properties.

Energetics of the B-H transition in supercoiled DNA carrying d(CT)x.d(AG)x and d(C)n.d(G)n inserts.

We have studied the B-H transition in the d(AG)x inserts of varying length under superhelical stress. The new data and previously published results for the d(G)31 insert are treated within a phenomenological model of the B-H transition, making it possible to obtain, for the first time, the energy parameters of the B-H transition in the d(AG)x and d(G)n sequences.

[Inclusion of biotinylated analogs of dUTP and dCTP in DNA by DNA-polymerases. Cloning DNA fragments, containing biotinylated deoxyribouridine in E. coli].

The synthesis of a biotinylated derivative of dCTP, viz. N4-[(N-biotinyl)-4-amino-butoxyl]-2'-deoxycytidine 5'-triphosphate (I), is described. DNA polymerase I (Klenow fragment) incorporates (I) in DNA chains instead of thymidine, although with a lower efficiency than previously described biotinylated dUTP derivative (II), whereas highly purified DNA polymerase alpha from human placenta uses as substrate derivative (II) but not (I).

[Localization of a lysine residue near the site of initiating substrate binding of T7 bacteriophage RNA polymerase].

A highly selective affinity label was introduced into the T7 phage RNA polymerase by means of GMP ortho-formylphenyl ester and [alpha-32P]UTP nearby the enzyme's active site, which was located using limited cleavage technique. Hydroxylamine, bromine, N-chlorosuccinimide, and cyanogen bromide were employed as the reagents. Analysis of gel-electrophoretic patterns of the cleavage products led to a conclusion that Lys631 is the target of labelling.

[Rapid automated synthesis of polydeoxynucleotides].

A rapid automatic method of synthesis of deoxypolynucleotides from 5'-O-dimethoxytritylnucleoside-3'-H-phosphonates is described. An improved construction of synthesizer "Gene-2" adapted for this method has been developed. The modified scheme of synthesis included detritylation with trifluoroacetic acids in dichloromethane, washing with acetonitrile instead of pyridine--acetonitrile mixture and one-step oxidation with iodine solution in acetic acid and pyridine instead of two-step oxidation in the presence of amines.

[An improved chemico-enzymatic method of synthesizing long DNA segments].

A simple and economy method of the biochemical assembling of long double-stranded DNA segments is described. A single-stranded polydeoxynucleotide 122 bases long representing a fragment of synthetic gene of human beta-interferon was assembled from three synthetic fragments 36 (two) and 50 bases long on four complementary 12-mers as templates. This single-stranded polynucleotide was converted, in the presence of DNA polymerase 1 and a 12-meric primer, in to the full-length double-stranded DNA (the beta-interferon gene segment).

The energetics of the B-Z transition in DNA.

The paper deals with the energetics of the transition to left-handed Z form in DNA with an arbitrary base sequence. There is a brief outline of the statistical-mechanical model of the B-Z transition allowing for three possible states of each base pair. The parameters of the model can be determined by comparing the theory with experimental data for the B-Z transition in inserts with given sequences in circular DNA.

[Current approaches to the synthesis and reconstruction of genetic material].

Advanced approaches to the synthesis and reconstruction of genetic material developed in the Institutes of Molecular Biology and Genetics during the past years are summarized. The evolution of methods for oligonucleotide synthesis and scopes for their use in gene production are discussed. The principles of localised mutagenesis methods developed in the Institute are described, such as: a) mutagenesis directed to the regulatory gene regions; b) segment-localized mutagenesis; c) mutagenesis directed by phosphotriester analogues of oligonucleotides.

[Sequences homologous to the rat ID element in mouse DNA].

Sequences homologous to the rat frain specific identifier sequences were found in the mouse genome. These sequences were defined by dot- and blotting-hybridisation in different DNA and RNA preparations. It was shown that there are (1-2) X 10(3) copies of these sequences per genome.

Cyclic GMP phosphodiesterase from cattle retina. Amino acid sequence of the gamma-subunit and nucleotide sequence of the corresponding cDNA.

The primary structure of the gamma-subunit of cyclic GMP phosphodiesterase was determined by parallel analysis of the amino acid sequence of the protein and nucleotide sequence of the corresponding cDNA. The enzyme gamma-subunit contains 87 amino acid residues, its N-terminal amino group being acetylated.

[Plasmid vector pSSC1 for cloning synthetic polynucleotides and their regeneration with a unique nucleotide sequence].

For subcloning separate synthetic gene fragments, a plasmid vector pSSC1 was constructed by inserting a synthetic polylinker into plasmid pBR 327 at the EcoRI-PstI sites. There are two FokI and HgaI sites at the ends of this polylinker in the opposite orientation, with the EcoRI and PstI sites between them. DNA fragments cloned at the EcoRI and PstI sites can be regenerated by either FokI or HgaI, the EcoRI and PstI sites being deleted from the cloned sequences.