Daoyu Profile picture
9 Jun, 20 tweets, 7 min read
nature.com/articles/s4159…
The most dishonest paper known.
The paper in fact, compared only icSARS-CoV Urbani, which is in fact the only coronavirus RGS that made use of the bgII restriction enzyme. All current
nature.com/articles/s4159…
ncbi.nlm.nih.gov/pmc/articles/P…
sciencedirect.com/science/articl…
RGS systems for SARS-CoV-2 made use of the type IIS enzyme BsaI and Esp3I/BsmBI. None have used that Urbani BgII because the genome is too different to be compatible with that old system.
dN introduced for these specific sites likely disrupted key function and consequently type IIS enzymes was used ncbi.nlm.nih.gov/pmc/articles/P… for all subsequent RGS systems for Coronaviruses.

link.springer.com/protocol/10.10…
In deed, all subsequent infectious clone systems used on Coronaviruses are based on type IIS enzymes ncbi.nlm.nih.gov/pmc/articles/P… not type IIG enzymes as in SARS-CoV Urbani.
They refused to accept the fact that basically no current papers that published infectious SARS-CoV-2 clones have ever made the clone to be compatible with SARS-CoV Urbani.

Considering these modern systems, SARS-CoV-2 is the only one that is clonable in this manner.
zenodo.org/record/4772451…
In fact, their use of MP789 indicating that it is in fact a clone—the virus never existed in the samples while the animals were still alive,
but only entered the samples afterward as contamination first and deliberate spike-in after. “Viral replication” without ACE2 expression when the S have been found only to use ACE2? Fake samples smooshed together with animal sample as matrix.
Notice how the S is swapped. BsaI/BsmBI, not BgII. ImageImageImage
“„The original fragments E and F were shortened to leave spike gene as an independent fragment. The new fragments were designated as Es and Fs. BsaI or BsmBI sites were introduced into the junctions of Es/Spike and Spike/Fs.
Then any spike could be substituted into the genome of SARSr-CoV WIV1 through this strategy.“”
ncbi.nlm.nih.gov/pmc/articles/P…

sciencedirect.com/science/articl…
BsmBI/BsaI type IIS no seem’s is THE earliest mean to clone a Coronavirus. BgII based ligation is only used for Urbani.
sciencedirect.com/science/articl…
Even SARS-Urbani have been cloned without that BgII. Image

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More from @Daoyu15

11 Jun
@Rossana38510044 @halvorz @ydeigin @stuartjdneil @brianholtz1965 @notoriousFIL The selection aren’t purifying, for starters. RaTG13 arm carried loads and loads of unnatural mutations comprising about 1/3 the total divergence from RaTg13 to SARS-CoV-2. The most parsimonious explanation for the large number of changes for MN996532 is by intentional tampering.
@Rossana38510044 @halvorz @ydeigin @stuartjdneil @brianholtz1965 @notoriousFIL Much of the changes are simulated. See article on simulation of large phylogenetic divergence and rescue of live virus based on simulated genomes. pubmed.ncbi.nlm.nih.gov/19036930/ see supplementary methods. The majority of SARS-CoV-2 nucleotides that differ from RaTg13 agrees with
Read 34 tweets
9 Jun
@InWuchang @amymaxmen There exist no need for any hacker to hack a scientific database about viruses (entirely unprofitable to hack and have no significance pre-outbreak whatsoever) at September 2019. The database went offline before the need to hack arises. And it have continued to be down
@InWuchang @amymaxmen Even until today. web.archive.org/web/2020012520… the database was stuck in an infinite loop and inaccessible even today. No hacking attempts are persistent over a year and a half. The database is not an “excel spreadsheet” as proclaimed by Shi in her interview. It is a 61.5MB SQL
Read 28 tweets
8 Jun
@lab_leak @uacjess CGGCGG is banned because it is an extremely powerful immune adjuvant. Feline coronavirus have CGGCGA precisely because CGGCGG is too immunogenic and will destroy the ability for it to persist in nature.
@lab_leak @uacjess CCACGGCGAGCA more precisely. Notice that this break up the palindrome within the sequence—CpG palindrome sequences are considered one of the best vaccine adjuvants known for mRNA vaccination. The CGGCGG itself is specifically patented as part of the adjuvant sequence.
@lab_leak @uacjess journals.asm.org/doi/abs/10.112…
In fact, it is so crippling that the ZAP protein, normally only able to restrict CpG-rich viral genomes, restricts SARS-CoV-2 just as efficiently as any other CoVs. The S gene expression is specifically restricted.
Read 15 tweets
7 Jun
@NeBirgitta @Parsifaler The extreme specificity to the lungs at the expense of other organs (especially the intestines, which is the reservoir of CoVs in the wild) is an extremely unnatural property. Wild animals don’t typically contact through the respiratory route—contact between different wild
@NeBirgitta @Parsifaler Animals are mediated primarily through the fecal-oral route as feces are more durable than aerosols, which don’t stay for time sufficient to span between the average time between meeting between individuals of typical wild animals. WuHu-1, or D614, does not have efficient
@NeBirgitta @Parsifaler Transmission through aerosols (unlike G614). An infection that is focused on the lower respiratory tract is not really transmissive but is very severe and will kill the host rapidly. This is the polar opposite to what that is needed for persistent transmission and survival in
Read 8 tweets
1 Jun
SARS-CoV-2 Can’t infect a BAT! RpYN06 can NEVER meet that S!
We investigated the susceptibility of primary cells from Rhinolophus ferrumequinum and Myotis species, as well as of established and novel cell lines from Myotis myotis, Eptesicus serotinus, Tadarida brasiliensis and Nyctalus noctula, to SARS-CoV-2 infection. None of these cells
were sensitive to infection, not even the ones expressing detectable levels of angiotensin-converting enzyme 2 (ACE2), which serves as the viral receptor in many mammalian species. The resistance to infection was overcome by expression of human ACE2 (hACE2) in three cell lines,
Read 6 tweets

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