A New Generation of Animal Cell Expression Vectors Based on the Semliki Forest Virus Replicon

1991 ◽  
Vol 9 (12) ◽  
pp. 1356-1361 ◽  
Author(s):  
Peter Liljeström ◽  
Henrik Garoff
1988 ◽  
Vol 103 (3-4) ◽  
pp. 157-166 ◽  
Author(s):  
A. Jalanko ◽  
A. Kallio ◽  
I. Ulmanen

2017 ◽  
Author(s):  
M. Butler ◽  
U. Reichl

Author(s):  
Robert E. Kingston ◽  
Randal J. Kaufman ◽  
C.R. Bebbington ◽  
M.R. Rolfe

2021 ◽  
Vol 15 (5) ◽  
pp. e0009374
Author(s):  
Kit Man Chai ◽  
Tsai-Teng Tzeng ◽  
Kuan-Yin Shen ◽  
Hung-Chun Liao ◽  
Jhe-Jhih Lin ◽  
...  

The development of efficient vaccines against COVID-19 is an emergent need for global public health. The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major target for the COVID-19 vaccine. To quickly respond to the outbreak of the SARS-CoV-2 pandemic, a nucleic acid-based vaccine is a novel option, beyond the traditional inactivated virus vaccine or recombinant protein vaccine. Here, we report a DNA vaccine containing the spike gene for delivery via electroporation. The spike genes of SARS-CoV and SARS-CoV-2 were codon optimized for mammalian cell expression and then cloned into mammalian cell expression vectors, called pSARS-S and pSARS2-S, respectively. Spike protein expression was confirmed by immunoblotting after transient expression in HEK293T cells. After immunization, sera were collected for antigen-specific antibody and neutralizing antibody titer analyses. We found that both pSARS-S and pSARS2-S immunization induced similar levels of antibodies against S2 of SARS-CoV-2. In contrast, only pSARS2-S immunization induced antibodies against the receptor-binding domain of SARS-CoV-2. We further found that pSARS2-S immunization, but not pSARS-S immunization, could induce very high titers of neutralizing antibodies against SARS-CoV-2. We further analyzed SARS-CoV-2 S protein-specific T cell responses and found that the immune responses were biased toward Th1. Importantly, pSARS2-S immunization in hamsters could induce protective immunity against SARS-CoV-2 challenge in vivo. These data suggest that DNA vaccination could be a promising approach for protecting against COVID-19.


Author(s):  
Veronika Altmannova ◽  
Andreas Blaha ◽  
Susanne Astrinidis ◽  
Heidi Reichle ◽  
John R. Weir

The successful production of recombinant protein for biochemical, biophysical and structural biological studies critically depends on the correct expression organism. Currently the most commonly used expression organisms for structural studies are E. coli (ca. 70% of all PDB structures) and the baculovirus/ insect cell expression system (ca. 5% of all PDB structures). While insect cell expression is frequently successful for large eukaryotic proteins, it is relatively expensive and time consuming compared to E. coli expression. Frequently the decision to carry out a baculovirus project means restarting cloning from scratch. Here we describe an integrated system that allows the simultaneous cloning into E. coli and baculovirus expression vectors using the same PCR products. The system offers a flexible array of N- and C-terminal affinity, solublisation and utility tags, and the speed allows expression screening to be completed in E. coli, before carrying out time and cost intensive experiments in baculovirus. Importantly, we describe a means of rapidly generating polycistronic bacterial constructs based on the hugely successful biGBac system, making InteBac of particular interest for researchers working on recombinant protein complexes.


BioTechniques ◽  
1997 ◽  
Vol 23 (3) ◽  
pp. 402-407 ◽  
Author(s):  
M.I. Cockett ◽  
R. Ochalski ◽  
K. Benwell ◽  
R. Franco ◽  
J. Wardwell-Swanson

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