Selective n-butanol production by Clostridium sp. MTButOH1365 during continuous synthesis gas fermentation due to expression of synthetic thiolase, 3-hydroxy butyryl-CoA dehydrogenase, crotonase, butyryl-CoA dehydrogenase, butyraldehyde dehydrogenase, and NAD-dependent butanol dehydrogenase.

Acetogen Clostridum sp. MT1962 produced 287 mM acetate (p < 0.005) and 293 mM ethanol (p < 0.

"Curing" of plasmid DNA in acetogen using microwave or applying an electric pulse improves cell growth and metabolite production as compared to the plasmid-harboring strain.

Plasmid-free acetogen Clostridium sp. MT962 electrotransformed with a small cryptic plasmid pMT351 was used to develop time- and cost-effective methods for plasmid elimination. Elimination of pMT351 restored production of acetate and ethanol to the levels of the plasmid-free strain with no dry cell weight changes.

Cre-lox66/lox71-based elimination of phosphotransacetylase or acetaldehyde dehydrogenase shifted carbon flux in acetogen rendering selective overproduction of ethanol or acetate.

Acetogen strain Clostridium sp. MT1121 produced 300 mM acetate (p<0.005) and 321 mM ethanol (p<0.

Selective production of acetone during continuous synthesis gas fermentation by engineered biocatalyst Clostridium sp. MAceT113.

Aims: To engineer acetogen biocatalyst capable of fermenting synthesis gas blend to acetone as the only liquid carbonaceous product.

Methods And Results: The metabolic engineering comprised inactivation of phosphotransacetylase via integration of a cassette comprising synthetic genes erm(B), thiolase and HMG-CoA synthase. Acetaldehyde dehydrogenase was inactivated via integration of a cassette consisting of synthetic genes cat, HMG-CoA lyase and acetoacetate decarboxylase.

Elimination of acetate production to improve ethanol yield during continuous synthesis gas fermentation by engineered biocatalyst Clostridium sp. MTEtOH550.

Acetogen strain Clostridum sp. MT653 produced acetate 273 mM (p < 0.005) and ethanol 250 mM (p < 0.

[The preparative method for 2-fluoroadenosine synthesis].

The preparative method for the synthesis of 2-fluoroadenosine starting from commercially available guanosine was developed. It included the intermediate formation of 2-amino-6-azido-9-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)purine, which was isolated exclusively in the tetrazolo[5,1-i]-form {5-amino-7-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)-7H-tetrazolo[5,1-i]purine}. The latter compound was converted by the Schiemann reaction to 6-azido-2-fluoro-9-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)purine, which was isolated at an 80% yield after careful optimization of the process.

[15-19-mer synthetic oligoribonucleotide as a model of the translation initiation segment of the phage fr replicase gene].

Automatic solid-phase synthesis of 15-19-meric oligoribonucleotides was carried out using 5'-O-dimethoxytrityl-2'-O-(2-tetrahydropyranyl)- [or 2'-O-(2-tetrahydrofuranyl)-] N-acylribonucleoside-3'-O-(methyl-N, N-diisopropyl)phosphoramidite synthons and 1-H-tetrazole as an activator. Comparative analysis of the template activity of the oligoribonucleotides synthesized, which are models of the translation initiation region of the replicase gene of MS2 and fr phages, showed that the minimal active fragment of RNA is a 16-mer containing the initiation AUG codon of the gene, a short spacer, a Shine-Dalgarno domain, and the 5'-terminal AUGA sequence with a functionally important termination AUG codon.

[The characteristics of the accumulation of metals in the hair of children].

Gender- and colour-associated quantitative features of accumulation of zinc, copper, nickel, lead, molybdenum and chromium have been studied in the hair of children aged 3-4 yr residing in different districts of Kiev. Inverse relationship was found between accumulation of lead and zinc, zinc and copper; direct proportionality between copper and lead, chromium and molibdenum. The levels of nickel detected in 34% of the hair samples did not permit establishing the pattern of this metal relations to other metallic elements.

Complete nucleotide sequence of the group I RNA bacteriophage fr.

We report the complete nucleotide sequence of the group I RNA bacteriophage fr. The entire genome consists of 3575 nucleotides, six nucleotides more than the only other sequenced group I representative, MS2. The greatest divergence between these phages occurs in the 5' terminal region of the A gene, while the lysis-replicase gene overlap, the coat gene and the central region of the replicase gene are highly conserved.

Affinity labelling of Escherichia coli ribosomes with a benzylidene derivative of AUGU6 within initiation and pretranslocational complexes.

Affinity labelling of E. coli ribosomes with the 2',3'-O-[4-(N-2-chloroethyl)-N-methylamino]benzylidene derivative of AUGU6 was studied within the initiation complex (complex I) obtained by using fMet-tRNAMetf and initiation factors and within the pretranslocational complex (complex II) obtained by treatment of complex I with the ternary complex Phe-tRNAPhe.GTP.

[Primary structure of a fragment of cDNA from phage fr].

The nucleotide sequence of a 1392 bp fragment of phage fr cDNA has been determined. The fragment contains 3'-terminal part of the A-protein gene, the complete coat protein gene, and beginning of the replicase gene. A comparison between the sequences of the corresponding genes and regulatory regions from the phage fr and MS2 genomes reveals 320 base changes.

[Study of the mRNA-binding region of ribosomes at different steps of translation. II. Affinity modification of Escherichia coli ribosomes by benzylidene derivative of AUGU6 in the 70S initiation complex].

2',3'-O-(4-[N-(2-chloroethyl)-N-methylamino]) benzylidene derivative of AUGU6 was used for identification of the proteins in the region of the mRNA-binding centre of E. coli ribosomes. This derivative alkylated ribosomes (preferentially 30S ribosomal) with high efficiency within the 70S initiation complex.

Novel human leukocyte interferon subtype and structural comparison of alpha interferon genes.

In the cDNA library of virus-induced human leukocytes a novel subtype of IFN-alpha gene has been identified and sequenced, named IFN-alpha-N. A comparison of nucleotide sequences within the genes coding for different subtypes of human leukocyte interferon (IFN-alpha) has revealed the natural hybrid structure of individual alpha-IFNs-H,B,F, and N. Certain regions of the genes for IFN-alpha-H,B,F,N show a homology with one of the two structurally distinct groups of leukocyte interferons--either IFN-alpha-A,D or IFN-alpha-C,C1 utilized as reference standards.

[Study of the ribosome mRNA-binding region during different stages of translation. I. Functional activity of mRNA analogs, AUGU6 and its benzylidene derivatives, in ribosome-dependent protein synthesis].

Hexaribouridylic acid, prepared by digestion of poly(U) with cobra venom endonuclease, and trinucleotide AUG synthesized chemically by triester approach were joined by RNA-ligase to yield a nonaribonucleotide AUGU6 bearing the initiation codon at its 5'-terminus. 2',3'-O-(4-[N-(2-chloro(or hydroxy) ethyl-N-Methylamino])- benzylidene residues were introduced at the 3'-terminus of oligonucleotide AUGU6 and its benzylidene derivatives AUGU6CHRCl or AUGU6CHROH were obtained. The mRNA analogs synthesized were tested for their template activity in the formation of 70S initiation complex.

The regulatory region of phage fr replicase cistron. III. Initiation activity of specific fr RNA fragments.

RNA fragments from phage fr covering the complete or part of the replicase cistron initiation region have been used as templates in the formation of a ribosomal initiation complex in vitro. The results so obtained together with our earlier findings in a similar approach applied to fragments of the structurally related RNA from phage MS2 have allowed us to pinpoint the boundaries of the replicase cistron initiation region on phage RNA. A structural model of the above initiation region has been provided which shows that besides the minimal initiation region (comprises the Shine-Dalgarno sequence and initiator AUG), the flanking regions are also involved and are responsible for additional interactions with the ribosome.

[Regulatory region of fr phage replicase cistron. II. Isolation and structure of specific fr RNA fragments].

A new set of short RNA templates has been prepared for functional studies in initiation of translation in vitro. Number of individual RNA fragments which contain complete or part of the initiatory region of phage fr replicase cistron were isolated from complex fr RNA--fr coat protein. Their primary structure were determined by using standard fingerprint technique and rapid gel sequencing.

[Preparation of RNAase-free E. coli ribosomes active in initiation of protein biosynthesis].

The level of contaminating RNAases in the main components of the protein biosynthesis initiation system, the initiation factors and ribosomes of E. coli, was studied. It was shown that the ribosomes are the major source of contaminating RNAases.