排序方式: 共有7条查询结果,搜索用时 0 毫秒
1
1.
Silaeva Yu. Yu. Kirikovich Yu. K. Skuratovskaya L. N. Deikin A. V. 《Russian Journal of Developmental Biology》2018,49(6):356-361
Russian Journal of Developmental Biology - The technology of creating genetically modified animals (placental mammals) by microinjection into the pronucleus of a fertilized egg suggests, as one of... 相似文献
2.
Yaroslav G. Gurskiy David G. Garbuz Nataliya V. Soshnikova Aleksey N. Krasnov Alexei Deikin Vladimir F. Lazarev Dmitry Sverchinskyi Boris A. Margulis Olga G. Zatsepina Vadim L. Karpov Svetlana N. Belzhelarskaya Evgenia Feoktistova Sofia G. Georgieva Michael B. Evgen’ev 《Cell stress & chaperones》2016,21(6):1055-1064
The production of major human heat shock protein Hsp70 (HSPA1A) in a eukaryotic expression system is needed for testing and possible medical applications. In this study, transgenic mice were produced containing wild-type human Hsp70 allele in the vector providing expression in the milk. The results indicated that human Hsp70 was readily expressed in the transgenic animals but did not apparently preserve its intact structure and, hence, it was not possible to purify the protein using conventional isolation techniques. It was suggested that the protein underwent glycosylation in the process of expression, and this quite common modification for proteins expressed in the milk complicated its isolation. To check this possibility, we mutated all presumptive sites of glycosylation and tested the properties of the resulting modified Hsp70 expressed in E. coli. The investigation demonstrated that the modified protein exhibited all beneficial properties of the wild-type Hsp70 and was even superior to the latter for a few parameters. Based on these results, a transgenic mouse strain was obtained which expressed the modified Hsp70 in milk and which was easy to isolate using ATP columns. Therefore, the developed construct can be explored in various bioreactors for reliable manufacture of high quality, uniform, and reproducible human Hsp70 for possible medical applications including neurodegenerative diseases and cancer. 相似文献
3.
Yu. Yu. Silaeva A. A. Kalinina M. S. Vagida L. M. Khromykh A. V. Deikin T. G. Ermolkevich E. R. Sadchikova I. L. Goldman D. B. Kazansky 《Biochemistry. Biokhimii?a》2013,78(5):549-559
Peripheral T lymphocytes can be subdivided into naive and antigen-experienced T cells. The latter, in turn, are represented by effector and central memory cells that are identified by different profiles of activation markers expression, such as CD44 and CD62L in mice. These markers determine different traffic of T lymphocytes in the organism, but hardly reproduce real antigenic experience of a T lymphocyte. Mechanisms of homeostasis maintenance of T lymphocytes with different activation phenotypes remain largely unknown. To investigate impact of T cell receptor (TCR) transgenic chains on formation of T lymphocytes, their peripheral survival and activation surface phenotypes, we have generated the transgenic mouse strain expressing transgenic β-chain of TCR 1D1 (belonging to the Vβ6 family) on the genetic background B10.D2(R101). Intrathymic development of T cells in these transgenic mice is not impaired. The repertoire of peripheral T lymphocytes in these mice contains 70–80% of T cells expressing transgenic β-chain and 20–30% of T cells expressing endogenous β-chains. The ratio of peripheral CD4+CD8? and CD4?CD8+ T lymphocytes remained unchanged in the transgenic animals, but the percent of T lymphocytes with the “naive” phenotype CD44?CD62L+ was significantly increased, whereas the levels of effector memory CD44+CD62L? and central memory CD44+CD62L+ T lymphocytes were markedly decreased in both subpopulations. On the contrary, T lymphocytes expressing endogenous β-chains had surface phenotype of activated T cells CD44+. Thus, for the first time we have shown that the pool of T lymphocytes with different activation phenotypes depends on the structure of T cell receptors. 相似文献
4.
5.
R. V. Zvartsev D. S. Korshunova E. A. Gorshkova M. A. Nosenko K. V. Korneev O. G. Maksimenko I. V. Korobko D. V. Kuprash M. S. Drutskaya S. A. Nedospasov A. V. Deikin 《Doklady. Biochemistry and biophysics》2018,483(1):344-347
To model human interleukin-6 (hIL-6) associated diseases, unique mice with transgenic overexpression of human IL-6 and reporter fluorescent protein EGFP in cells of macrophage-monocyte lineage were generated using loxP–Cre system. High level of hIL-6 production by macrophages and monocytes, as confirmed in vitro in primary culture of bone marrow-derived macrophages, in vivo resulted in early postnatal death in vivo, presumably, due to the effect of overexpression of hIL-6 on hematopoiesis. 相似文献
6.
Yaroslav Gursky Robert Bibilashvili Mikchail Minashkin Alex Krasnov Alex Deikin Tatyana Ermolkevich Andrey Popov Lilia Verbovaya Nicolai Rutkevich Alexsander Shevelev Sofia Georgieva Sergey V. Razin Igor Goldman Elena Sadchikova 《Transgenic research》2009,18(5):747-756
Human pro-urokinase expressed in the mammary glands of transgenic animals is quickly activated and converted to urokinase
by proteases that are present in the milk. Thus, it is nearly impossible to isolate full-sized pro-urokinase from the milk
of transgenic animals. To solve this problem, we constructed transgenic mice that express human pro-urokinase and modified
ecotin, which is a potent serine protease inhibitor from E. coli, in their mammary glands. The gene encoding ecotin was modified so as to enhance its specificity for the human urokinase-type
plasminogen activator. Co-expression of modified ecotin and human pro-urokinase in the mammary glands allows for purification
of full-length human pro-urokinase from these transgenic mice. The results described here suggest a general way of preventing
the activation of zymogens that are expressed in the mammary glands of transgenic animals by co-expression of a zymogen along
with a protease inhibitor. 相似文献
7.
E. S. Zvezdova Yu. Yu. Silaeva M. S. Vagida E. V. Maryukhnich A. V. Deikin T. G. Ermolkevich S. G. Kadulin E. R. Sadchikova I. L. Goldman D. B. Kazansky 《Molecular Biology》2010,44(2):277-286
Transgenic animal analysis has become a key approach used to study the gene functions and to model various human diseases,
including autoimmune disorders. Such disorders are caused by the activation of T-cell clones whose T-cell receptors (TCRs)
have a high affinity for syngeneic MHC molecules. The genes coding for the α and β chains of the autoreactive TCR were cloned
from hybridoma 7, which was specific for syngeneic Ab MHC class II molecules. Amplified DNA fragments containing rearranged
genomic DNA of the α and β chains of hybridoma 7 were cloned into special cassette vectors that contained the natural promoter
and enhancer elements ensuring direct expression of the α- and β-chain genes in T cells of transgenic animals. The animals
obtained with the vectors expressed the α or β chain on the majority of peripheral Tcells. The animals are suitable for studying
the features of the intrathymic selection and maturation of T cells and provide an experimental model for developing new approaches
to therapy of autoimmune diseases. 相似文献
1