New insights into stability of recombinant adenovirus vector genomes in mammalian cells

2012 ◽  
Vol 91 (1) ◽  
pp. 2-9 ◽  
Author(s):  
Christina Rauschhuber ◽  
Nadja Noske ◽  
Anja Ehrhardt
Keyword(s):  
Mammalian Cells ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Recombinant Adenovirus Vector

scholarly journals Expression of the human androgen receptor in eukaryotic cells using a recombinant adenovirus vector yields high levels of the soluble, functional receptor protein

2001 ◽  
Vol 27 (3) ◽  
pp. 321-328
Author(s):  
S Zoppi ◽  
DR Allman ◽  
RD Gerard ◽  
MJ McPhaul
Keyword(s):  
Androgen Receptor ◽  
Mammalian Cells ◽  
Large Scale ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Receptor Protein ◽  
Eukaryotic Cells ◽  
Native Form ◽  
Recombinant Adenovirus Vector ◽  
High Level

The androgen receptor (AR) and closely related members of the steroid receptor family have proven difficult to obtain in native form in large quantities. In the case of the human AR (hAR), high-level expression in prokaryotic or non-mammalian cells leads to the synthesis of a high proportion of non-binding, insoluble, or degraded forms of the receptor protein. To circumvent these difficulties, we have constructed a recombinant adenovirus that directs the expression of hAR under the control of a potent, constitutive promoter. Infection of eukaryotic cells with this recombinant virus leads to the synthesis of large quantities of the intact AR. In contrast to expression methods designed to direct the full-length AR in bacteria, yeast, and insect cells, AR expressed in mammalian cells using this adenoviral vector accumulates at high levels, retains many properties of the native AR, and is not rapidly proteolyzed. This method will prove useful for large-scale preparations of hAR for use in functional and structural studies.

scholarly journals The Himar1 mariner transposase cloned in a recombinant adenovirus vector is functional in mammalian cells

1998 ◽  
Vol 26 (16) ◽  
pp. 3687-3693 ◽  
Author(s):  
L. Zhang ◽  
U. Sankar ◽  
D. J. Lampe ◽  
H. M. Robertson ◽  
F. L. Graham
Keyword(s):  
Mammalian Cells ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Recombinant Adenovirus Vector

Development of a recombinant adenovirus vector production system free of replication-competent adenovirus by utilizing a packaging size limit of the viral genome

2011 ◽  
Vol 158 (1-2) ◽  
pp. 154-160 ◽  
Author(s):  
Takayuki Suzuki ◽  
Tomomi Sasaki ◽  
Koyori Yano ◽  
Fuminori Sakurai ◽  
Kenji Kawabata ◽  
...  
Keyword(s):  
Production System ◽  
Viral Genome ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Size Limit ◽  
Recombinant Adenovirus Vector

A simplified cloning strategy for the generation of an endothelial cell selective recombinant adenovirus vector

2006 ◽  
Vol 135 (1) ◽  
pp. 127-135 ◽  
Author(s):  
Chitladda Mahanivong ◽  
Jörg A. Krüger ◽  
Dafang Bian ◽  
Ralph A. Reisfeld ◽  
Shuang Huang
Keyword(s):  
Endothelial Cell ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Cloning Strategy ◽  
Recombinant Adenovirus Vector

Regulation of N-myc gene expression: use of an adenovirus vector to demonstrate posttranscriptional control

1990 ◽  
Vol 10 (12) ◽  
pp. 6700-6708
Author(s):  
L E Babiss ◽  
J M Friedman
Keyword(s):  
Recombinant Adenovirus ◽  
Wilms Tumor ◽  
Fetal Brain ◽  
Adenovirus Vector ◽  
Cell Types ◽  
Adult Brain ◽  
Posttranscriptional Control ◽  
Equivalent Rate ◽  
Myc Gene ◽  
Recombinant Adenovirus Vector

We present evidence that differences in the levels of N-myc mRNA among different cell types are the result of posttranscriptional control. First, we noted that while steady-state mouse N-myc mRNA could be detected only in fetal mouse brain, it was transcribed at an equivalent rate in adult brain, liver, spleen, and placenta and in fetal brain. Similarly, the human N-myc gene was transcribed at an equivalent rate in HeLa cells, which do not accumulate this RNA in the cytoplasm, and cell lines G401 (a Wilms tumor-derived cell line) and SKNMc (established from a primitive neuroepithelioma), which do express N-myc RNA. As expected, the N-myc promoter functioned at equivalent rates, as demonstrated by the level of a reporter gene, when introduced into these cell types by using a recombinant adenovirus vector. The suggestion that posttranscriptional mechanisms control the level of this RNA was supported by the observation that sequences in the N-myc third exon specifically decreased the level of E1A mRNA when these sequences were placed downstream of the E1A promoter in a recombinant adenovirus. Finally, we further localized these sequences to a 600-bp fragment of the third exon by introducing various subclones of this sequence downstream of the E1A promoter in both viral and plasmid vectors.

Expression of human α1-antitrypsin using a recombinant adenovirus vector

FEBS Letters ◽  
1990 ◽  
Vol 267 (1) ◽  
pp. 60-62 ◽  
Author(s):  
P. Gilardi ◽  
M. Courtney ◽  
A. Pavirani ◽  
M. Perricaudet
Keyword(s):  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Recombinant Adenovirus Vector

Experimental Study on Construction of Human Hiwi Adenovirus Vector and Apoptosis of K562 Cells Induced by Transfected Vector

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4721-4721
Author(s):  
Xiaoyan Dong ◽  
Yanfang Jiang ◽  
Mengmeng Liu ◽  
Wei Li ◽  
Ziling Liu
Keyword(s):  
Stem Cells ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
K562 Cells ◽  
Leukemia Stem Cells ◽  
Positive Clone ◽  
Coding Region ◽  
Overlap Extension ◽  
Recombinant Adenovirus Vector ◽  
Pcr Method

Abstract Abstract 4721 Background: Leukemia stem cells are responsible for the genesis, progression, drug resistance and relapse of leukemia. Hiwi gene is an important divisive regulatory factor, which plays a role in maintaining the resting stage and down-regulating the cell cycle of stem/progenitor cells. We acquired human overall Hiwi coding region genes and constructed an adenovirus vector carrying human Hiwi with fluorescin. This study will not only establish the foundation of the further study in function and mechanism of Hiwi induce the differentiation and apoptosis of leukemia stem cells, but also provide the theoretical basis for searching new therapeutic target and method for leukemia. Methods: Using the overlap extension PCR method to amplify overall Hiwi coding region genes and insert overall Hiwi coding region genes into Flag-IRES –hrGFP vector that carrying green fluorescin with Gateway clone technology to construct pDown-Hiwi-3×flag-IRES-hrGFP. After transform the vector to Stb13, we screen the positive clone with PCR method, extract the plasmid and process recombination reaction between pDown-Hiwi-3×flag-IRES-hrGFP clone vector and pAV.Des1d expression vector to abtain the pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant. Screening the positive clone with PCR and extracting the plasmid which were digested by enzyme and then examined by sequencing consequences. At last packaged Ad-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant is obtained. Transfect the recombinant adenovirus vector to K562 cells, and then identify the function of the cells. Results: Successfully clone human overall Hiwi coding region genes with the technology of overlap extension PCR and construct the adenovirus vector recombinant with enzyme cutting identification and gene sequencing examination. Conclusions: Successfully construct Ad-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant and establish the foundation for the further study of Hiwi gene in leukemia stem cells. The transfected recombinant adenovirus vector can induce K562 cells to apoptosis. [U1]ɾ3ý£z Disclosures: No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document