1. The Yeast is Red and Many are Dead Thread
Sub Yeast Thread 1
2. via @ydeigin and @franciscodeasis
it’s a very disturbing paper if its findings are verified: not only SARS2 and RaTG13 have signs of genetic manipulation in yeast but SARS1 too.
WIV is known to have eyed yeast reverse genetic systems with interest (see attached WIV thesis
3. The Author has published quite a few interesting papers with 1870 citations.
scholar.google.com/citations?user…
One on the Association between influenza vaccination rates and SARS-CoV-2 outbreak infection rates in OECD countries
papers.ssrn.com/sol3/papers.cf…
4. Random Yeast Papers - WIV
Construction and Rescue of a Functional Synthetic Baculovirus
pubs.acs.org/doi/10.1021/ac…
5. Honours in Yeast (08/2020) - Yeast Virus Cookbook
Every Tweet is one Tab Less ;)
In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline
honours.eu/publications/i…
and pdf:
ncbi.nlm.nih.gov/pmc/articles/P…
6. Yeast containing SARS-COV- 2
“The exciting thing about the yeast is that . . . it’s fast,” says expert Susan Weiss
Scientists Clone SARS-CoV-2 Genome with Quick Yeast-Based Method (May 2020)
the-scientist.com/news-opinion/s…
7. Dr. Weiss's Magic Yeast

What this means is that instead of having to reconstruct the viral DNA first before introducing it into cells, you instead “put all these fragments in the yeast and it magically puts them together,” says Weiss.
8. TAR, YAC & DNA

transformation associated recombination (TAR)

yeast artificial chromosome (YAC)

Data Not Available (DNA) Don't believe me? Ask WIV!
9. Amazing Recombination Capacity

“The capacity to recombine things to a YAC by themselves is just amazing,” says Jores.
10. In Vitro Ligation

Refers to this UTMB paper:

An Infectious cDNA Clone of SARS-CoV-2

cell.com/cell-host-micr…

Note: Pei Yong Shi and James Le Duc
11. Method from above UTMB paper

An Infectious cDNA Clone of SARS-CoV-2

cell.com/cell-host-micr…
12. One Week Is All It Takes

Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform

nature.com/articles/s4158…
13. Nice Clear Layman's Explanation (Thank God)

Researchers develop synthetic novel coronavirus clones

phys.org/news/2020-05-s…
14. Well illustrated background reading

Billy Recommends: More Yeast with your Whining!

Synthetic genome engineering forging new frontiers for wine yeast

tandfonline.com/doi/full/10.10…
15. Virus Like Particle Vaccine Manufacture from Yeast
(Illustrated Master's Thesis) 2020
digitalcommons.mtu.edu/cgi/viewconten…
16. Synthetic viruses—Anything new?

asks Volker Thiel in 2018

The generation of synthetic poxviruses as described by Evans and colleagues and cloning of synthetic DNA using TAR in yeast are illustrated

journals.plos.org/plospathogens/…
17. Yeast-based synthetic genomics platform to genetically reconstruct RNA viruses exist.

How much were they used by Chinese Researchers in the years leading up to the pandemic is an interesting research project.
18. WIV Research (2018) - Yeast

Advances and challenges in enveloped virus-like particle (VLP)-based vaccines

Shiyu Dai, Hualin Wang, Fei Deng

State Key laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan

immunologyresearchjournal.com/public/article…
19. Time for a break!

Unroll the Yeast is Red @threadreaderapp
20. Revenge of Yeast
a WIV Thesis that @TheSeeker268 found recently is by Lei Ping Zeng (2017) & it mentions WIV's interest in yeast recombination methods

Reverse Genetic System of Bat SARS-like Coronaviruses & Function of ORFX
drive.google.com/drive/folders/…
21. Zeng Translated
(by @TheSeeker268 & @franciscodeasis )
docs.google.com/document/d/1uZ…
Only 2 references to yeast recombination but obviously on their radar back in 2017.
22. Across ocean & back in time to Wizard of UNC

Thanks Jim Haslam @jhas5

Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV

journals.plos.org/plosone/articl…
23. I am getting so sick of Yeast!
I will just dump yeast infected papers here for others to vomit over
Yeast for virus research (2017) Richard Yuqi Zhao
ncbi.nlm.nih.gov/pmc/articles/P…
24. Enough Yeast!
2007: Saccharomyces cerevisiae: a versatile eukaryotic system in virology
…robialcellfactories.biomedcentral.com/articles/10.11…
2010: A High-Throughput Approach for Studying Virus Replication in Yeast
…rentprotocols.onlinelibrary.wiley.com/doi/10.1002/97…
25. I will finish with yeast until i get more yeastinspiration

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More from @BillyBostickson

Aug 22
1. Nord Stream Update

3 years for truth to emerge

@MerlynThomas on 27/09/22 for @BBCWorld

Ukraine accuses Russia of pipeline terror attack

bbc.com/news/world-eur…

@JamesCrisp6 on 28/09/22 for @Telegraph

Why Putin would want to blow up Nord Stream 2

archive.ph/JCtYB x.com/Panchenko_X/st…
2. Hilariously

Merlyn Thomas has been working for the "BBC disinformation unit" since September 2021.

He certainly earned his UK STATE propaganda pay check with Nord Stream!

3. Invitation to venture down a Baltic rabbit hole!

You are hereby invited to read my 682 tweet thread investigating all angles of the Nord Stream sabotage, which ends here:



Thanks @Erkperk @VasilijProzorov @JeffreyBrodsky5 @MichaKobs @bopanc @liscer72
Read 4 tweets
Aug 13
Baric's hACE2 transgenic mice were shared with the WIV for GOF like virus experiments designed to "assess" the potential for coronaviruses to jump from animals to humans.

We asked how many were sent to WIV

researchgate.net/publication/35…

Record via @jhas5 here

arlo.riseforanimals.org/entity/univers… x.com/jhas5/status/1…
2. Baric spills his humanized mice beans to Father Racaniello during a heart rending TWIV confessional in March 2020, admitting he sent his hACE2 mice to WIV in 2018 (as part of the infamous NIAID grant mentioned above):



Timestamp; 01:00:30 - 01:03:15microbe.tv/twiv/twiv-591/
3. Daszak

"Our R01 work shows that bats in South China harbor many SARSr-CoVs (50+), some of which are able to use human ACE2 to enter human cells, infecting humanized mouse models to cause SARS-like illness, and evade available therapies or vaccines"

ecohealthalliance.org/wp-content/upl…
Read 9 tweets
Aug 11
1. New England Biolabs & Novagen

The amplified Rp3-S1 fragment was digested with BamHI & XhoI & ligated into the expression vector pMAL-C2X (New England Biolabs).

The 5 truncated fragments were digested with BamHI & XhoI and ligated into the expression vector pET32a (Novagen).
2. Promega & Takara

Protein expression & purification

After purification, recombinant plasmid was transformed into BL21 competent cells (purchased from Promega)

Recovered PCR product & expression vector pET32a were digested with restriction endonucleases BamHI & XhoI (Takara).
3. Takara (2)

After DNA recovery, the PCR product was ligated to the expression vector pET32a using T4 DNA ligase (Takara)
Read 32 tweets
Jul 26
@breakfast_dogs @gadboit @NLink247 1. Insect cell baculovirus cell cultures become contaminated with insect genomic material.
2. Replication process can lead to presence of viral DNA & cellular components in culture medium.
3. This LEADS to contamination of viral product with baculovirus & insect cell-derived DNA.
@breakfast_dogs @gadboit @NLink247 Contamination

4. Expression of SC-2 Spike RBD Using the Insect Cell-Baculovirus System

pmc.ncbi.nlm.nih.gov/articles/PMC97…

5. Co-expression vs. co-infection using baculovirus expression vectors in insect cell culture

pmc.ncbi.nlm.nih.gov/articles/PMC71…

6. insect cell lines
sciencedirect.com/science/articl…
@breakfast_dogs @gadboit @NLink247 Did WIV use insect cell systems with SARS-C0V-2?

7. Insect cell baculovirus expression systems

x.com/BillyBostickso…

8. DENG Fei, Chief Investigator in Research Group on Viruses Resources at WIV engineered baculovirus-insect cell expression systems.

buzzchronicles.com/b/all/17652/
Read 13 tweets
Jul 22
1. Kadlec awaiting confirmation for his new role

2. Kadlec's report references pangolins 76 times

bush.tamu.edu/wp-content/upl…

3. Time to read my pangolin coronavirus report?

researchgate.net/publication/39…

4. DRASTIC research cited
@gdemaneuf @ydeigin @BillyBostickson @C_Small_ x.com/BoDeepest/stat…
1. the article linked is quite good, more comments and analysis will be posted here soon

city-journal.org/article/robert…Image
2. Possible lab manipulations

"2) insertion of a pangolin virus recombinant spike or the spike’s RBD protein that includes a sequence for an FCS and an integrin-binding protein" Image
Read 76 tweets
Jul 9
Pressure is needed on people like @realannapaulina @RepLuna & @GOPoversight team as their mandate may dissolve in August.

They did Kennedy, but they are supposed to do UFO files & CoVid-19 origins before they dissolve.

We also need to know about the subpoenas sent by @RandPaul
If you are a concerned US citizen please directly contact the people listed here:

oversight.house.gov/subcommittee/t…
Read 8 tweets

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