https://www.biorxiv.org/content/10.1101/2021.01.27.427998v1
Abstract
We engineered three SARS-CoV-2 viruses containing key spike mutations from the newly emerged United Kingdom (UK) and South African (SA) variants: N501Y from UK and SA; 69/70-deletion+N501Y+D614G from UK; and E484K+N501Y+D614G from SA. Neutralization geometric mean titers (GMTs) of twenty BTN162b2 vaccine-elicited human sera against the three mutant viruses were 0.81- to 1.46-fold of the GMTs against parental virus, indicating small effects of these mutations on neutralization by sera elicited by two BNT162b2 doses.
（以We開頭，得多麽自信自豪啊！我們做了，服不服？）
在正文方法一節中，作者寫到了怎麽做出這三個變異株並在Vero細胞傳代增殖的。這充分說明所合成改造過的病毒具有生物活性，不是偽（模擬）病毒。
Three recombinant SARS-CoV-2 mutants (N501Y,
102
Δ
69/70-N501Y+D614G, E484K+N501Y+D614G in spike protein) were prepared on the genetic
103
background of an infectious cDNA clone derived from clinical strain WA1 (2019-
104
nCoV/USA_WA1/2020)
5
by following the PCR-based mutagenesis protocol as reported
105
previously
7
. The full-length infectious cDNAs were
in vitro
ligated and used as templates to
106
transcribe full-length viral RNA. Mutant viruses (P0) were recovered on day 2 from Vero E6 cells
107
after electroporation of the
in vitro
RNA transcripts. P1 viruses were harvested as stocks by
108
passaging the P0 virus once on Vero E6 cells. The titers of P1 viruses were determined by
109
plaque assay on Vero E6 cells. The genome sequences of the P1 viruses were validated by
110
Sanger sequencing. The detailed protocol was recently reported
17
.
The full-length infectious cDNAs were
in vitro
ligated and used as templates to
106
transcribe full-length viral RNA. Mutant viruses (P0) were recovered on day 2 from Vero E6 cells
107
after electroporation of the
in vitro
RNA transcripts. P1 viruses were harvested as stocks by
108
passaging the P0 virus once on Vero E6 cells. The titers of P1 viruses were determined by
109
plaque assay on Vero E6 cells. The genome sequences of the P1 viruses were validated by
110
Sanger sequencing. The detailed protocol was recently reported
17
.
The full-length infectious cDNAs were
in vitro
ligated and used as templates to
106
transcribe full-length viral RNA. Mutant viruses (P0) were recovered on day 2 from Vero E6 cells
107
after electroporation of the
in vitro
RNA transcripts. P1 viruses were harvested as stocks by
108
passaging the P0 virus once on Vero E6 cells. The titers of P1 viruses were determined by
109
plaque assay on Vero E6 cells. The genome sequences of the P1 viruses were validated by
110
Sanger sequencing. The detailed protocol was recently reported
17
.”
