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EF1Ξ±-Promotor
ββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββ
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EF1Ξ±-Promotor bezeichnet einen Promotor des Elongationsfaktors EF1Ξ±. Er ist neben dem CAG-Promotor und dem CMV-Promotor einer der meistverwendeten Promotoren in Expressionsvektoren.
Contents
β’ Eigenschaften
β’ StΓΆrfaktoren
β’ Literatur
β’ Einzelnachweise
ββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββββ
Eigenschaften
Der humane EF1Ξ±-Promotor steuert in Zellen die konstitutive Genexpression des Elongationsfaktors 1Ξ±. Er erzeugt dabei im Vergleich zu vielen anderen Promotoren der Wirtszelle grΓΆΓere Mengen an Proteinen. Dadurch kΓΆnnen dem Promotor gentechnisch nachgeschaltete Transgene verstΓ€rkt abgelesen werden, wodurch mehr mRNA und in Folge mehr rekombinante Proteine erzeugt werden. Daher wird er zur Γberexpression verschiedener rekombinanter Proteine in Zellkulturen von SΓ€ugetier-Zelllinien oder -PrimΓ€rzellen, darunter in humanen T-Zellencite-ref-pmid31014302-1-0[1] und NK-Zellen,cite-ref-pmid30871576-2-0[2] CHO-Zellen,cite-ref-pmid28161546-3-0[3] Rattenzellen,cite-ref-pmid26769799-4-0[4] Rinderfibroblasten,cite-ref-pmid36441701-5-0[5] Schweinefibroblastencite-ref-pmid23379578-6-0[6] und Schizosaccharomyces pombe, verwendet.cite-ref-pmid18437702-7-0[7] Der essentielle Bereich des EF1Ξ±-Promotors ist mit etwa 210 Basenpaaren einer der kΓΌrzesten starken Promotoren fΓΌr Expressionsvektoren.cite-ref-pmid37198545-8-0[8]
ZelltypabhΓ€ngigkeit
Der EF1Ξ±-Promotor zΓ€hlt mit dem CAG-Promotor zu den Promotoren mit gleichmΓ€Γig starker Expression in verschiedenen SΓ€ugetierzellen.cite-ref-qin-9-0[9]
StΓΆrfaktoren
Die AktivitΓ€t des EF1a-Promotors kann in transgenen Tieren aufgrund umfangreicher DNA-Methylierung reprimiert werden (Silencing). Daher ist er zumindest fΓΌr die forcierte Expression einiger Proteine, wie das Beispiel ACE2 zeigt, nicht geeignet.cite-ref-pmid35960480-10-0[10]
Literatur
β’ D. W. Kim, T. Uetsuki, Y. Kaziro, N. Yamaguchi, S. Sugano: Use of the human elongation factor 1 alpha promoter as a versatile and efficient expression system. In: Gene. Band 91, Nummer 2, Juli 1990, S. 217β223, PMID 2210382.
Einzelnachweise
cite-note-pmid31014302-11. β Y. Lv, F. J. Xiao, Y. Wang, X. H. Zou, H. Wang, H. Y. Wang, L. S. Wang, Z. Z. Lu: Efficient gene transfer into T lymphocytes by fiber-modified human adenovirus 5. In: BMC biotechnology. Band 19, Nummer 1, April 2019, S. 23, doi:10.1186/s12896-019-0514-x, PMID 31014302, PMC 6480437 (freier Volltext).
cite-note-pmid30871576-22. β S. V. Kulemzin, D. A. Matvienko, A. H. Sabirov, A. M. Sokratyan, D. S. Chernikova, T. N. Belovezhets, A. N. Chikaev, A. V. Taranin, A. A. Gorchakov: Design and analysis of stably integrated reporters for inducible transgene expression in human T cells and CAR NK-cell lines. In: BMC medical genomics. Band 12, Suppl 2MΓ€rz 2019, S. 44, doi:10.1186/s12920-019-0489-4, PMID 30871576, PMC 6417161 (freier Volltext).
cite-note-pmid28161546-33. β M. D. Rocha-PizaΓ±a, G. Ascencio-Favela, B. M. Soto-GarcΓa, M. L. Martinez-Fierro, M. M. Alvarez: Evaluation of changes in promoters, use of UCOES and chain order to improve the antibody production in CHO cells. In: Protein expression and purification. Band 132, April 2017, S. 108β115, doi:10.1016/j.pep.2017.01.014, PMID 28161546.
cite-note-pmid26769799-44. β A. I. Garcia Diaz, B. Moyon, P. M. Coan, N. Alfazema, L. Venda, K. Woollard, T. Aitman: New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein. In: Disease models & mechanisms. Band 9, Nummer 4, April 2016, S. 463β471, doi:10.1242/dmm.024208, PMID 26769799, PMC 4852507 (freier Volltext).
cite-note-pmid23379578-66. β S. Petkov, P. Hyttel, H. Niemann: The choice of expression vector promoter is an important factor in the reprogramming of porcine fibroblasts into induced pluripotent cells. In: Cellular reprogramming. Band 15, Nummer 1, Februar 2013, S. 1β8, doi:10.1089/cell.2012.0053, PMID 23379578.
cite-note-pmid37198545-88. β J. Li, Q. Liang, H. Zhou, M. Zhou, H. Huang: Profiling the impact of the promoters on CRISPR-Cas12a system in human cells. In: Cellular & molecular biology letters. Band 28, Nummer 1, Mai 2023, S. 41, doi:10.1186/s11658-023-00454-9, PMID 37198545, PMC 1019003 (freier Volltext).
cite-note-qin-99. β J. Y. Qin, L. Zhang, K. L. Clift, I. Hulur, A. P. Xiang, B. Z. Ren, B. T. Lahn: Systematic comparison of constitutive promoters and the doxycycline-inducible promoter. In: PLOS ONE. Band 5, Nummer 5, Mai 2010, S. e10611, doi:10.1371/journal.pone.0010611, PMID 20485554, PMC 2868906 (freier Volltext).
cite-note-pmid35960480-1010. β N. Battulin, A. Korablev, A. Ryzhkova, A. Smirnov, E. Kabirova, A. Khabarova, T. Lagunov, I. Serova, O. Serov: The human EF1a promoter does not provide expression of the transgene in mice. In: Transgenic research. Band 31, Nummer 4β5, Oktober 2022, S. 525β535, doi:10.1007/s11248-022-00319-5, PMID 35960480, PMC 9372930 (freier Volltext).