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单基因缺失重组仙台病毒载体的高效增殖

Efficient propagation of single gene deleted recombinant Sendai virus vectors.

作者信息

Bernloehr Christian, Bossow Sascha, Ungerechts Guy, Armeanu Sorin, Neubert Wolfgang J, Lauer Ulrich M, Bitzer Michael

机构信息

Internal Medicine I, University Clinic Tübingen, D-72076, Tübingen, Germany.

出版信息

Virus Res. 2004 Feb;99(2):193-7. doi: 10.1016/j.virusres.2003.11.005.

Abstract

Recombinant Sendai virus vectors (SeVV) have become an attractive tool for basic virological as well as for gene transfer studies. However, to (i) reduce the cellular injury induced by basic recombinant SeV vectors (encoding all six SeV genes as being present in SeV wild-type (wt) genomes) and to (ii) improve SeV vector safety, deletions of viral genes are necessary for the construction of superior SeVV generations. As a strong expression system recombinant replication-incompetent adenoviruses, coding for SeV proteins hemagglutinin-neuraminidase (HN), fusion (F), or matrix (M), were generated and successfully employed for the propagation of single gene deleted (DeltaHN, DeltaF, DeltaM) recombinant SeVV. Further investigations of the propagation procedures required for single gene deleted recombinant SeVV demonstrated (i) modifications of the cell culture medium composition as well as (ii) incubation with vitamin E as crucial steps for the enhancement of SeVV-DeltaHN, -DeltaF, or -DeltaM viral particle yield. Such optimized propagation procedures even led to a successful propagation of HN-deleted viral particles (SeVV-DeltaHN), which has not been reported before.

摘要

重组仙台病毒载体(SeVV)已成为基础病毒学研究以及基因转移研究的一种有吸引力的工具。然而,为了(i)减少基础重组SeV载体(编码SeV野生型(wt)基因组中存在的所有六个SeV基因)诱导的细胞损伤,以及(ii)提高SeV载体的安全性,构建更优质的SeVV代次时需要删除病毒基因。作为一种强大的表达系统,编码SeV蛋白血凝素神经氨酸酶(HN)、融合蛋白(F)或基质蛋白(M)的重组无复制能力腺病毒被构建出来,并成功用于传播单基因缺失(ΔHN、ΔF、ΔM)的重组SeVV。对单基因缺失重组SeVV所需传播程序的进一步研究表明,(i)细胞培养基成分的改变以及(ii)与维生素E一起孵育是提高SeVV-ΔHN、-ΔF或-ΔM病毒颗粒产量的关键步骤。这种优化的传播程序甚至成功实现了缺失HN的病毒颗粒(SeVV-ΔHN)的传播,这在此前尚未见报道。

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