[Structural organization of C-hordein genes from barley]. / Strukturnaia organizatsiia genov C-gordeinov iachmenia.

Three nucleotide sequences from the barley genome, containing the C-hordein genes lambda CH1, lambda CH3, and lambda CH5, were cloned. They were shown to have different physical maps, and the structural organization of homologous sequences was found to be highly conservative. The surrounding sequences of C-hordein genes contain insertions specific in length and copy number. A Hind III fragment of about 3000 bp in size of the lambda CH5 clone contains a highly repeated nucleotide sequence. The lambda CH3 clone contains two "head-to-head" repeated C-hordein genes. This is the first evidence that hordein genes may be located in the immediate vicinity of each other in a 17,800-bp region. A 919-bp nucleotide sequence of one of the C-hordein genes from this clone, pCHOR3, was determined. An open reading frame of 363 bp codes for a potential polypeptide consisting of 121 amino acids. The main part of the mature polypeptide consists of repeated octapeptide Pro-Gln-Gln-Pro-Phe-Pro-Gln-Gln motifs. The coding region has no introns. The 5' untranslated region contains regulatory loci specific for prolamin genes: the -300 element and the CAAT, AGGA, and TATA boxes. Significant differences in the lengths of the coding regions of cloned genes were observed: 363 and 1000 bp in the lambda CH3 clone, 500 bp in the lambda CH1 clone, and 600 bp in the lambda CH5 clone, which may be a molecular cause of C-hordein size polymorphism.

[Nucleotide sequence of the barley C-hordein gene]. / Nukleotidnaia posledovatel'nost' gena C-gordeina iachmenia.

The nucleotide sequence of barley C-hordein gene lambda CH4 and its flanking regions of 2820 bp length was determined. The gene contains no introns and codes for 310 amino acid long polypeptide. The 94% of the deduced amino acid sequence of the mature protein (291 amino acids) is made up of a repeating octapeptide motiff, PQQPEPQQ, which is repeated throughout the peptide chain between a unique 12 amino acid long NH2 terminal and a unique 6 amino acid long COOH-terminal end. In the 5' non-coding region there are TATA-, AGGA-, CAAT-and "endosperm" boxes. The 3' non-coding region has two polyadenylation signals. Compared with the published C-hordein sequences, our gene contains a number of insertions, deletions and substitutions. In the 3'-untranslated region there are two insertions 157 and 23 bp long. For the longer insertion no significant homology was found in the Gene Bank datebase. This insertion is the largest known rearrangement in the otherwise highly conservative surroundings of barley storage protein genes.

[The genomic organization and localization on the chromosomes of the HvRT family of DNA repetitive sequences in barley]. / Genomnaia organizatsiia i lokalizatsiia na khromosomakh HvRT-semeistva povtoriaiushchikhsia posledovatel'nostei DNK iachmenia.

HvRT family of repetitive DNA sequences from barley genome appears to have complex hierarchical organization. Tandem repetition of 118-bp monomers constitutes lower level of HvRT-family organization. Amplification units of the higher level consist of several contiguous 118-bp monomers. RFLP between different species and cultivars of barley resulted from the differences in the higher-order repeat structure. Individual chromosomes of barley contain specific HvRT subfamilies. This family also possesses separate domains differing in the restriction enzyme sites density. HvRT family is presented in the genomes of H. vulgare, H. leporinum, H. murinum, H. jubatum, but is absent in the genomes of H. marinum, H. geniculatum and wheat.

[Cloning and partial characteristics of the barley D-hordein sequence]. / Klonirovanie i chastichnaia kharakteristika posledovatel'nosti D-gorgeina iachmenia.

A number of clones containing major endosperm-specifically transcribed gene copies were selected from a cDNA library developed on the basis of barley endosperm mRNA. Approx. 30% of the recombinant clones carried sequences homologous to mRNA of various cereal storage proteins. Some of them appeared to be related to cDNA clones of wheat and barley storage proteins. The typical insert length ranged from 0.3 to 1.7 kB. A couple of clones among them were selected which revealed positive hybridization with all probes used. The positive signals disappeared after stringent washing of the filters. The nucleotide sequences of two representatives of the group were determined and corresponding amino acid sequence deduced after subsequent computer analysis. The comparison with known cereal storage protein genes revealed relatively high homology level with the central part of wheat high molecular weight (HMW) glutenine subunit genes. The fact suggests the cloned gene to belong to barley D-hordein family.

[The cloning of "residual" DNA and the determination of the primary structure for the fragments of the barley-specific family of tandem repetitive sequences]. / Klonirovanie "ostatochnoi" DNK i opredelenie pervichnoi struktury fragmentov iachmen'-spetsificheskogo semeistva tandemno povtoriaiushchikhsia posledovatel'nostei.

High molecular weight "relic" DNA fraction can be separated from the bulk of barley DNA digested with different restriction enzymes by agarose gel electrophoresis. The majority of AluI-relic DNA clones contained barley simple sequence satellite DNA and other families of repetitive DNA. The clones representing HvRT family were sequenced. This family was found to be barley-specific. It is composed of tandemly arranged 118-bp monomers and presents in 7 x 10(5) copies in barley genome.

[Nucleotide sequence of the B1-hordein gene of barley Hordeum vulgare L]. / Nukleotidnaia posledovatel'nost' gena B1-gordeina iachmenia Hordeum vulgare L.

The gene encoding B1 hordein of Hordeum vulgare (cv. Donetsky 4) was cloned and entirely sequenced. It contains no introns and codes for of 293 amino acids long polypeptide with molecular weight 33418. Our clone differs from the previously sequenced B1 hordein genes in some positions within the coding region (there are 4 nucleotide changes and a 12 bp deletion, as compared with the pB11 cDNA clone, and 5 nucleotide changes, as compared with the pBHP184 genomic clone). These changes result in polymorphism of amino acid sequences at 5 positions. 5'-flanking region contains putative regulatory and promoter sequences and differs from that of the pBHP184 clone in 3 positions.

[Isolation of DNA fragments of the barley genome, supporting autonomous regulation of plasmids in Saccharomyces cerevisiae]. / Vydelenie fragmentov DNK genoma iachmenia, podderzhivaiushchikh avtonomnuiu replikatsiiu plazmid v drozhzhakh-sakharomitsetakh.

BamHI fragments of the barley genomic DNA were cloned in Escherichia coli cells on the vector plasmid YIp5 carrying the URA3 gene of Saccharomyces cerevisiae. Yeast cells were transformed by individual plasmid DNA preparations from each clone selected. Approximately 10% of the studied plasmids are able to replicate in yeast cells. Five barley DNA fragments which could support replication of recombinant plasmids in yeast cells belong to moderately repeated DNA and are dispersed through the chromosomes.

[Organization and primary nucleotide sequence of 5S rRNA genes in barley Hordeum vulgare]. / Organizatsiia i pervichnaia nukleotidnaia posledovatel'nost' genov 5S pRNK u iachmenia (Hordeum vulgare).

A BamHI DNA fragment of 301 bp corresponding to the main repeating unit of 5S rRNA was isolated from barley genomic DNA. The primary nucleotide sequence of this fragment was determined and a high level of homology was found between coding sequences of 5S rRNA genes of barley, wheat and rye. At the same time, spacer's nucleotide sequences of different species of cereals were changed dramatically. At least two types of 5S rRNA tandem repeats of 301 and 450 bp were found in barley genome. Polymorphism for restriction fragment length in 5S rRNA repeats allowed to discriminate between all barley varieties used in this work.

[A new class of mobile genetic elements of Drosophila melanogaster]. / Novyi klass mobil'nykh geneticheskikh élementov Drosophila melanogaster.

Clone Dm A89 was obtained upon cloning of DNA fragments coding abundant poly(A+)RNA's of D. melanogaster. Dm A89 was identified as a new transposable element using in situ hybridization with polytene chromosomes of two independent highly isogenic lines of D. melanogaster oregon RC and gt wa Dm A89 hybridizes with approximately 20 sites in each line. A portion of Dm A89 is homologous to the distal part of type I ribosomal gene insertion sequence and is highly repetitive. Two other sections of the clone have much less redundancy. The unity of the three fragments is not casual, as revealed by cloning of some other genomic sequences homologous to Dm A89. Dm A89 is actively transcribed throughout the development of D. melanogaster and produces polyadenylated RNA 1.1 kb long.

[Effect of diucifon and its structural precursors on inflammatory and pain reactions]. / Vliianie diutsifona i ego strukturnykh predshestvennikov na vospalitel'nuiu i bolevuiu reaktsii.

The anti-inflammatory and analgesic action of diucifon, methyluracil and 4,4'-diaminodiphenylsulfone (DDS) was studied in comparison with some nonsteroid preparations. In three traditional models of agar, kaolin and carrageenan paw edema and in the models of analgesia (convulsions, induced by intraabdominal administration of acetic acid to mice and hyperalgesia according to Randall-Selitto's test in rats), diucifon proved more active than its precursors. In Randall-Selitto's test the efficiency of diucifon was 1.5 times less than that of butadione but 24 times higher than that of methyluracil; DDS had no analgetic activity. Diucifon, methyluracil and DDS did not exert any ulcerogenic action.

[Visualization of elementary structures in polytene chromosomes of Drosophila melanogaster]. / Vizualizatsiia élementarnykh struktur v politennykh khromosomakh Drosophila melanogaster.

Electron microscopic study of total polytene chromosome preparations makes it possible to reveal elementary structures of polytene chromosomes. Elementary chromosomes are detected which contain chromomeric and interchromomeric regions. Transcription is seen in puffs and in many bands. Chromatin spreading shows that both chromomeric and interchromomeric regions of elementary chromosomes consist of nucleosomes. A ribbon-like model of polytene chromosome structure is suggested and discussed. In the ribbon-like chromosome each elementary chromosome gels conjugated only with the two adjacent sister chromosomes. Conjugation occurs only in the chromomeric regions, interchromomeric regions do not conjugate. Both chromomeric and interchromomeric regions are duplicated during polytenization which ensure proportional replication of all the bands and interbands in the course of polytenization.

[Molecular approaches to the systematic mapping of the human genome]. / Molekuliarnye podkhody k sistematicheskomu kartirovaniiu genoma cheloveka.

A review of current progress in human gene mapping methods is presented. The advantages and restrictions of several mapping methods are discussed. The main bulk of the review is concerned with perspectives of using special collection of "molecular-genetic markers" (MGM). These are cloned nucleotide sequences which allow to find population (and family) polymorphisms in three structural characteristics of homologous DNA fragments: in length, in regional chromosome location, in the number of local copies. Accordingly, three groups of MGM are analysed: (i) unique genome fragments, (ii) mobile dispersed genes and (iii) clustered DNA repeats. As the authors suggest, the second one has to be the most available tool for recognition of mother and father partners in all chromosome pairs. It is an important step to the progress in mapping not only monogenic traits but polygenic characters too, including multifactorial diseases in man.

[Transcription of portions of chromosomes corresponding to regions of intercalary heterochromatin in Drosophila melanogaster cell cultures]. / Transkriptsiia uchastkov khromosom, sootvetstvuiushchikh raionam interkaliarnogo geterokhromatina v kul'ture kletok Drosophila melanogaster.

Labelled RNA preparations (total newly synthesized RNA, as well as stable cytoplasmic RNA) isolated from a cell culture of Drosophila melanogaster were hybridized in situ with polytene chromosomes. Apart from the nucleolus, in all cases the regions adjacent to he chromocentre in the polytene chromosomes and the intercalary heterochromatin regions in the X chromosome and the autosomes are the most intensively labelled. In the case of asynapsis of polytene chromosomes in heterozygotes the label is detected in a number of intercalary heterochromatin sites in one homologous only ("the asymmetrical label"). The same kind of radioactivity distribution in intercalary heterochromatin regions was observed after a hybridization of polytene chromosomes with cloned DNA fragments (Ananiev et al.,, 1978, 1979) coding for the abundant classes of messenger RNA (Ilyin et al., 1978) in cultured D. melanogaster cells. In some regions of intercalary heterochromatin which do not contain these fragments the "asymmetrical" type of labelled distribution is observed after hybridization with cell RNA. These results lead one to regard the intercalary heterochromatin regions as "nests" comprising different types of actively transcribable genes, the composition of each nest varying in different stocks of D. melanogaster.

A new type of organization of the genetic material in eukaryotes.

An investigation of the properties of a number of genes of Drosophila, obtained by cloning recombinant DNAs, led to the detection of a new type of organization of genetic material. It was found that a number of actively working structural genes of Drosophila are represented by a large number of copies, scattered over its chromosomes. Their localization in the chromosomes is variable, although they are always detected in regions of intercalary heterochromatin. The latter evidently is a site of accumulation of various multiple genes.

[Rate of DNA synthesis and the size of replication units in cell culture of Drosophila melanogaster]. / Opredelenie razmerov replikonov i skorosti replikatsii DNK v kletok Drosophila Melanogaster.

The rate of DNA replication and the size of replication units was examined by means of pulse-labeling with 3H-thymidine and DNA autoradiography in the chromsomes of Drosophila melanogaster synchronized cells culture in vitro. The center-to-center distance between adjacent labeled section of DNA fibers was taken for a size of replication unit, i.e. replicon. DNA synthesis was revealed to start simultaneously in most replicons. The rate of elongation of labeled sections of DNA fibers is about 25 mu per hour (75 kb per hour). Taking into consideration bidirectinal DNA replication the rate of DNA replication per growing point is 12.5 mu per hour (38 kb per hour). The mean size of replicon is about 60 mu (130 kb) in cell culture of Drosophila melanogaster and most of these replicons vary from 20 to 80 mu. Calculation suggests that one replicon may consist of six chromomeres each being known to be formed by a segment of DNA molecular 10 mu long on average.

[DNA content, replication and transcription rate in different site of the X-chromosome of Drosophila melanogaster]. / Soderzhanie DNK, skorost' replikatsii i intensivnost' transkriptsii v razlichny razlichnykh uchastkakh raiona 1A1-3S8 X-khromosomy Drosphyla melanogaster

Duration of replication period and relative content of DNA and chromosomal newly-synthesized RNA were determined in nine sites of the region 1A1-3C8 in the polytene X-chromosome of D. melanogaster. Determination of DNA content by cytophotometry of Feulgen stained preparations and by autoradiography after long-term [3H]thymidine labeling gave similar results for all nine sites. The smallest portion of the chromosome in which DNA content could be measured by cytophotometry was about 2-10 bands. The mean DNA content per one band measured separately in 25 sites of the 1A1-3C8 region containing 109 bands differs in 8 times. Calculations show that different chromomeres of elementary chromosome (chromatide) contained from 8 to 65 X 10(3) bases pairs. No additional DNA replication was shown to be involved in the process of 2B region puffing. The duration of replication period was found to correlate negatively with the level of the transcription rate in nine sites of the 1A1-3C8 region of the X-chromosome (r = -0.65). The transcription rate correlates negatively with the DNA content in the largest band for each of nine sites. No significant correlation was obtained between duration of replication period and DNA content in the largest band for each of nine regions.