[The immunogenic properties of a recombinant vaccinia virus with an incorporated DNA copy of the 26S RNA of the Venezuelan equine encephalomyelitis virus].

A recombinant strain of vaccinia virus (VR26) containing a DNA-copy of the subgenomic 26S RNA of Venezuelan equine encephalomyelitis virus (VEE) inserted into the coding region of thymidine kinase (TK) gene was produced. This subgenomic RNA contained the genes for all structural proteins of the VEE virus, the strain Trinidad donkey (TRD). VR26 effectively expressed VEE virus glycoproteins on the membranes of the infected cells.

[Effective synthesis and cloning of the human interleukin-2 gene and its analog: expression of the interleukin-2 gene in E. coli cells].

Artificial DNA fragments encoding human interleukin-2 (133 a.a.) and its analogue (deletion of 14 C-terminal a.

[Design of recombinant-stable plasmids of the pFH series].

By cloning synthetic oligonucleotides into pUC18 plasmid, pFH123--pFH127 plasmids have been constructed. Their polylinker area, along with sites of widely used restriction endonucleases, contains two pairs each of FokI and HgaI sites in the opposite orientation to provide subfragment with unique predetermined 5'-ends. Comparative stability of the new plasmids and their derivatives has been studied and compared with that of the earlier constructed pMB plasmids.

[Plasmids pMB123 and pMB124--vectors for obtaining subfragments of DNA with random "sticky" ends].

For preparing a DNA fragment with unique protruding ends, plasmid vectors pMB123 and pMB124 were constructed by inserting a synthetic polylinker into plasmid pUR222 at the EcoRI-PstI sites. The polylinker contains two FokI and HgaI sites at its ends in opposite orientation flanking a combination of SalGI, AccI, HindII, HindIII (the latter site is absent from pMB124) and BamHI sites. DNA fragment cloned at the SalGI and BamHI sites can be regenerated by either FokI or HgaI treatment, the SalGI and BamHI sites being deleted from the cloned sequence.

[Identification in vivo of promoter activity in the left site of the att region of bacteriophage lambda DNA].

The promoter-probing vector (pSK plasmid) was explored for cloning of the fragments from lambda cI857 and lambda b2 DNAs containing different regions of the att site. We have constructed all-tet fusions where the fusions are: 1) HindIII/BamHI-491 base pairs (b. p.

[Effect of promoter duplication in the synthetic interferon gene on the level of its expression].

The expression of the artificial gene for the human leukocyte interferon (IFN) synthesized by the chemo-enzymic method has been studied in Escherichia coli cells. The genetic constructions in which the IFN gene transcription is carried out from A2 and B promoters of T7 phage or from E. coli lacZ UV5 gene promoter have been obtained, based on pEMB101 plasmid.

[Plasmid vector for cloning, determination of nucleotide sequence and directed assembly of DNA fragments].

A plasmid vector pNIMB has been constructed (starting) from the pUR222 plasmid as a result of substitution of the polylinker containing restriction sites: PstI, SalGI, AccI, HindII, BamHI EcoRI and by other synthetic linkers with additional sites for HindIII and HgaI. Plasmid pNIMB does not differ from the parent one phenotypically. Compared to pUR222 the vector contains an additional site for cloning HindIII fragments of DNA and allows to clone SalGI/BamHI- and PstI/SalGI-fragments.

[Synthesis and cloning of a DNA fragment containing a probable site for eukaryotic mRNA binding to ribosome].

A series of oligonucleotides, including two polynucleotides of 33 bases long, were synthesized by a solid-phase phosphotriester method. Potassium salt of 3-nitro-1,2,4-triazole in the presence of 18-crown-6 ether was used as nucleophilic catalyst. The partly complementary polynucleotides were elongated by DNA-polymerase I (Klenow fragment) to the full duplex, which was digested with SalGI and was inserted into a plasmid pUR222.

[Isolation and characterization of a phage T7 DNA fragment containing promoter B active in vivo and in vitro].

The fragment of 124 base pairs (Alu-124) has been isolated from the phage T7 DNA and cloned in the plasmid pSK. The sequence of the fragment was determined and position of this fragment on the physical map of phage DNA was found. It has been found that Alu-124 fragment contained an active in vivo and in vitro promoter for E.

[Synthesis of a 33-member polynucleotide containing the "core" att site of phage lambda DNA and its cloning].

Two polynucleotides containing 33 monomeric units were synthesized by a solid-phase phosphotriester method. These polynucleotides form a duplex with protruding 5'-ends, which allows to clone the duplex in EcoRI site of a cloning vehicle. Each polynucleotide was purified by electrophoresis in polyacrylamide gel, and the duplex obtained was cloned in EcoRI site of pUR 222 plasmid DNA.

[Integrase of lambda phage inhibits the activity of promotor Patt under in vivo and in vitro conditions].

In vitro and in vitro transcription of the region of lambda phage DNA containing promoter Patt has been studied. Active transcription was observed in the vicinity of the att site and at the promoter Patt in vivo during the first minutes after infection. This transcription considerably decreases at the 1th minute, and stops completely at the 15th minute after infection.