Enhanced transgene expression in mammalian cells by recombinant baculovirus vector containing bovine papillomavirus type 1 replication elements

2013 ◽  
pp. n/a-n/a
Author(s):  
Binbin Li ◽  
Changwei Yu ◽  
Juan Yin ◽  
Jiang Zhong
Keyword(s):  
Mammalian Cells ◽  
Transgene Expression ◽  
Recombinant Baculovirus ◽  
Bovine Papillomavirus ◽  
Baculovirus Vector ◽  
Expression In Mammalian Cells

Titration of K ATP Channel Expression in Mammalian Cells Utilizing Recombinant Baculovirus Transduction

2002 ◽  
Vol 8 (2) ◽  
pp. 99-111 ◽  
Author(s):  
Jeffrey L. Pfohl ◽  
Jennings F. Worley ◽  
J. Patrick Condreay ◽  
Gang An ◽  
Christopher J. Apolito ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Recombinant Baculovirus ◽  
Channel Expression ◽  
Expression In Mammalian Cells

Selective enhancement of bovine papillomavirus type 1 DNA replication in Xenopus laevis eggs by the E6 gene product

1989 ◽  
Vol 9 (2) ◽  
pp. 406-414
Author(s):  
H Romanczuk ◽  
W M Wormington
Keyword(s):  
Dna Replication ◽  
Xenopus Laevis ◽  
Gene Product ◽  
Mammalian Cells ◽  
Xenopus Laevis Oocytes ◽  
Bovine Papillomavirus ◽  
In Vitro Synthesis ◽  
E6 Gene ◽  
E6 Protein

Genetic analyses of bovine papillomavirus type 1 (BPV-1) DNA in transformed mammalian cells have indicated that the E6 gene product is essential for the establishment and maintenance of a high plasmid copy number. In order to analyze the direct effect of the E6 protein on the replication of a BPV-1-derived plasmid, a cDNA containing the BPV-1 E6 open reading frame was subcloned into an SP6 vector for the in vitro synthesis of the corresponding mRNA. The SP6 E6 mRNA was injected into Xenopus laevis oocytes to determine the subcellular localization of the E6 gene product and to analyze the effect of the protein on BPV-1 DNA replication. SP6 E6 mRNA microinjected into stage VI oocytes was translated into a 15.5-kilodalton protein that was specifically immunoprecipitated by antibodies directed against the E6 gene product. The E6 protein preferentially accumulated in oocyte nuclei, a localization which is consistent with the replicative functions in which it has been implicated. The expression of E6 in replication-competent mature oocytes selectively enhanced the replication of a BPV-derived plasmid, indicating a direct role for this gene product in the control of BPV-1 DNA replication.

Adenoviral vector platform for transduction of constitutive and regulated tricistronic or triple-transcript transgene expression in mammalian cells and microtissues

2006 ◽  
Vol 8 (10) ◽  
pp. 1208-1222 ◽  
Author(s):  
Valeria Gonzalez-Nicolini ◽  
Carlota Diaz Sanchez-Bustamante ◽  
Shizuka Hartenbach ◽  
Martin Fussenegger
Keyword(s):  
Mammalian Cells ◽  
Transgene Expression ◽  
Adenoviral Vector ◽  
Expression In Mammalian Cells

scholarly journals The food additive vanillic acid controls transgene expression in mammalian cells and mice

2011 ◽  
Vol 40 (5) ◽  
pp. e37-e37 ◽  
Author(s):  
Marc Gitzinger ◽  
Christian Kemmer ◽  
David A. Fluri ◽  
Marie Daoud El-Baba ◽  
Wilfried Weber ◽  
...  
Keyword(s):  
Vanillic Acid ◽  
Mammalian Cells ◽  
Transgene Expression ◽  
Food Additive ◽  
Expression In Mammalian Cells

scholarly journals Versatile multi-transgene expression using improved BAC TG-EMBED toolkit, novel BAC episomes, and BAC-MAGIC

2019 ◽  
Author(s):  
Binhui Zhao ◽  
Pankaj Chaturvedi ◽  
David L. Zimmerman ◽  
Andrew S. Belmont
Keyword(s):  
Mammalian Cells ◽  
Transgene Expression ◽  
Large Fraction ◽  
Fine Tuning ◽  
Cmv Promoter ◽  
Chromosome Position ◽  
High Level ◽  
Genomic Regions ◽  
Expression In Mammalian Cells

ABSTRACTAchieving reproducible, stable, and high-level transgene expression in mammalian cells remains problematic. Previously, we attained copy-number-dependent, chromosome-position-independent expression of reporter minigenes by embedding them within a BAC containing the mouseMsh3-Dhfrlocus (DHFR BAC). Here we extend this “BAC TG-EMBED” approach. First, we report a toolkit of endogenous promoters capable of driving transgene expression over a 0.01-5 fold expression range relative to the CMV promoter, allowing fine-tuning of relative expression levels of multiple reporter genes expressed on a single BAC. Second, we show small variability in both the expression level and long-term expression stability of a reporter gene embedded in BACs containing either transcriptionally active or inactive genomic regions, making choice of BACs more flexible. Third, we describe an intriguing phenomenon in which BAC transgenes are maintained as episomes in a large fraction of stably selected clones. Finally, we demonstrate the utility of BAC TG-EMBED by simultaneously labeling three nuclear compartments in 94% of stable clones using a multi-reporter DHFR BAC, constructed with a combination of synthetic biology and BAC recombineering tools. Our extended BAC TG-EMBED method provides a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies.

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