[The effect of regulatory sequences of alphaS1-casein gene on the expression of the lacZ-gene in loach Misgurnus fossilis L. transgenic embryos].

Transient expression of recombinant plasmids carrying the lacZ gene under the control of either bovine alphaS1-casein gene tissue-specific promoter-enhancer region or highly homologous goat alphaS1-casein gene promoter-enhancer region with supplementary regulatory sequences of the goat gene were studied in Misgurnus fossilis L. loach embryos. It has been shown previously that the expression of the constructs carrying these regulatory elements in transgenic mice occurred primarily in the mammary glands.

[Secretion of biologically active human granulocyte colony-stimulating factor (G-CSF) in milk of transgenic mice].

Two constructs were devised, containing the full-length gene of the human granulocyte colony-stimulating factor (G-CSF) fused with the 5' and 3' flanking promoter sequences of bovine alpha-S1-casein gene. Both constructs contained a 1518-bp fragment that included exons 18 and 19 and 320 bp of the 3' flanking region of bovine gene @CSN1S1, but differed in size of the 5' flanking sequences, which were of 721 bp, and exon 1 in construct pGCm1 and 2001 bp and exon 1 and intron 1 in construct pGCm2. With both constructs, transgenic mice were produced.

[Study of the transfer of foreign genes into mussel Mytilus Galloprovincialis Lam. eggs by spermatozoa].

The possibility of transferring exogenous DNA into eggs by mussel Mytilus galloprovincialis Lam. sperms both with the use of certain methods of transfection and without them was studied. The efficacy of egg fertilization by sperms treated with foreign DNA and the development of larvae at early stages of embryogenesis were evaluated.

[The transfer of foreign DNA by rabbit spermatozoa into hamster eggs].

Capacitated rabbit sperm was incubated with pCMVlacZ plasmid and then used to fertilize hamster oocytes liberated from zona pellucida. After treatment with DNase I, these oocytes were examined for the presence of exogenous DNA by PCR. DNA of pCMVlacZ was not found in oocytes after simple incubation with male gametes.

[Expression of the CMV-lacZ- and RSV-lacZ-genes in transgenic fish and mouse embryos].

The bacterial gene for beta-galactosidase under the control of LTR from either human cytomegalovirus (CMV-lacZ) or the Rous sarcoma virus (RSV-lacZ) was injected into fertilized eggs of Misgurnus fossilis L. loaches and F1 hybrid mice (CBA x C57Bl). Expression of the CMV-lacZ was observed in almost 100% of the loach embryos and larvae for two months following the first day of embryonic development.

[Analysis of expression of the RSV-lacZ-gene in transgenic embryos of the loach Misgurnus fossilis L. during different variations of injection].

Several series of microinjection of the RSV-lacZ gene (the Escherichia coli beta-galactosidase gene under the control of the long terminal repeat of the Raus sarcoma virus) into fertilized mud loach eggs were carried out. The expression of the transgene in 3-5 day-old fry was shown to depend neither upon the stage of fry development at which the RSV-lacZ gene was introduced (early blastodisc, late blastodisc, 2-blastomere embryo) nor upon the region of transgene injection (blastodisc cytoplasm or egg yolk). Additionally, when injected into yolk, a smaller number of transgene expression points were observed and their distribution was confined to the surface of the yolk sac.

[Amplification of the integrated bovine growth hormone transgene using the polymerase chain reaction (PCR)].

Conditions of the PCR amplification of unique fragments of bovine growth hormone (bGH) were found that allow reliable identification of transgenic rabbits for this gene. An integrated construction containing a part of MT promoter of metallothionein gene and chromosomal bGH gene was amplified. Two of 46 newborn lambs were found to be transgenic for the bGH gene by using primers BG3 and BG4 for amplification of the gene bGH fragment and its following restriction with pVUII.

[The production and characteristics of transgenic mice expressing the surface protein gene of the human hepatitis B virus].

Transgenic mice were obtained that contained a gene of human hepatitis B surface antigen (HBsAg). The integrated HBsAg DNA sequences were inherited in the normal Mendelian fashion. 6 of 25 investigated transgenic mice expressed the HBsAg.

[Genetic transformation of somatic cells. XI. The autonomic replication of the cloned DNA of the bovine papilloma virus in NIH/3T3 mouse fibroblasts and the changes in the growth characteristics of the transformed cells].

Mouse fibroblasts NIH 3T3 were transfected with the plasmid pBPV (142-6) containing full genome of bovine papilloma virus 1, and focuses of morphological transformation were selected 2-3 weeks later. DNA molecules, containing BPV-1 sequences, were isolated from extrachromosomal fraction of transformed clones suggesting stable autonomous replication of BPV in 3T3 NIH cells. In some rescued plasmids deletions spanning E6, 7 genes of BPV were found.

[Genetic transformation of somatic cells. X. Morphological transformation and changes in the growth characteristics of NIH 3T3 cells administered the oncogene v-myc].

Transfection of the plasmid containing the cloned gag-myc part of retrovirus MC 29 into mouse NIH 3T3 cells induces focuses of morphological transformation. Isolated morphological transformants have a decreased dependence on serum growth factors, a higher saturation density in monolayer, and an increased cloning efficiency on the glass and in agar. Induced traits are stably inherited, and may constitute the direct consequence of stable maintenance and expression of transfected oncogens.