[The forensic medical identification of the members of the Demidov family].

The objective of the present study was to elucidate the genetic profiles of the biological materials taken from four graves in the Demidov family vault in order to establish kinship between its members. According to the archival documents, two graves contained the remains of Pyotr Grigor'evich Demidov, an adjutant-general for the emperor Aleksandr II, and his wife Elizaveta Nikolaevna Demidova (Bezobrazova). Also, it was supposed that two other graves contained the remains of Grigory Petrovich Demidov and Ekaterina Petrovna Demidova (married name princess Kudasheva), the son and the daughter of P.G. Demidov and E.N. Demidova. The bodies remained in the half-ruined crypt during approximately 150 years under conditions of enhanced humidity and seasonal temperature fluctuations which made their bone tissue virtually unsuitable for the genetic analysis. Genotyping was performed with the use of standard AmpF/STR Identifiler-TM and AmpF/STR Yfiler-TM kits ("Applied Biosystems", USA). As a result of the study, the skeletal remains of the boy from grave No2 were identified as actually belonging to the son of P.G. Demidov and E.N. Demidova with a probability of no less than 99.999999998%. whereas the girl buried in grave No4, was not the daughter of these parents.

[Establishing the presence of blood in stains for material evidence by using the luminescent blood test]. / Ob ustanovlenii nalichiia krovi v piatnakh na veshchestvennykh dokazatel'stvakh s pomoshch'iu liuminestsentnogo gemotesta.

Two methods for objective persuasive identification of blood stains on pieces of material evidence are offered. These methods are 1000 times more sensitive than other methods used in forensic medical practice. Computer processing of the results permits calculating the content of hemoglobin derivatives in the examined material in amounts as low as 5-100 ng/100 microliters extraction. Using these methods, it is possible to prove the presence of blood by rapid analysis directly at the site of the crime in washed traces and in traces formed long ago.

[Increase in the yield of recombinant proteins in Saccharomyces cerevisiae yeasts as a result of optimizing their culture conditions]. / Povyshenie vykhoda rekombinantnykh belkov v drozhzhakh Saccharomyces cerevisiae v rezul'tate optimizatsii uslovii ikh kul'tivirovaniia.

The optimal one-stage scheme of the cultivation of the recombinant yeast strains was developed. In these strains, the expression of the foreign gene is induced at the medium depletion by phosphorus. The scheme was tested for the producers of human interleukin-2 and bovine gamma-interferon. In all the strains, the yield of cell biomass and the production of the foreign proteins increased as compared to the standard two-stage cultivation. The use of the developed scheme resulted in the threefold increase in the yield of the target proteins and in the productivity of the biotechnological process.

[Isolation of a yeast vector marked by the mutation of multiple resistance to antibiotics]. / Poluchenie drozhzhevogo vektora, markirovannogo mutatsiei mnozhestvennoi ustoichivosti k antibiotikam.

The plasmid mutation AntR determining multiple resistance to antibiotics--tetracycline and cycloheximide in Saccharomyces cerevisiae was earlier obtained and genetically characterized. In this work we describe experiments on cytoduction and transformation, proving the localization of this mutation in the yeast 2 mu DNA. As a result of cotransformation of the sensitive cells carrying a double mutation in the gene LEU2 with the yeast vector marked by LEU2 and 2 mu DNA obtained from the yeast AntR mutant, the Leu+ AntR clones were selected. Though the primary co-transformans contain both plasmids in an unlinked state, we managed to get clones in which the markers AntR and LEU2 were linked. The putative recombinant molecules were cloned in Escherichia coli and then introduced into the yeast recipient cells, differing by the presence of the endogenous 2 mu DNA. Retransformation of cir0 cells results in the appearance of the clones in which LEU2 and AntR markers segregate together. Thus, the result of cotransformation and selection in vivo is that the mutation of multiple resistance was included into the yeast vector plasmid, presumably, in its 2 mu part.

[Construction of vectors for eukaryotes on the basis of fragments of mitochondrial DNA of animals]. / Konstruirovanie vektorov dlia éukariot na osnove fragmentov mitokhondrial'noi DNK zhivotnykh.

The Bam HI fragment of rat liver mtDNA was used for construction and analysis of the cloning vehicles for prokaryota and eukaryota. Transformation of mutant bacterial cells deficient in DNA polymerase I polA with recombinant pBR-mtBA DNA was shown previously. Now the transformation of bacterial cells with recombinant pmt Tn9 DNA was observed. The Bam HI-A fragment of animal mtDNA may be used as vehicle replicon not only in bacterial cells but also in yeasts. The recombinant molecule was constructed from plasmide DNA YJpI which contained yeast chromosomal LEU2 gene and Bam HI-A mtDNA fragment.