#Shi & her 2022 #China Patent: BALB/c mouse adapted strain of #SARS related coronavirus WIV1-RsSHC014S and application thereof..
Inventors: Shi Zhengli, Lin Haofeng, Liu Meiqin, Jiang Rendi
Zhou Peng. #WIV1 #RsSHC014S
Inventors: Shi Zhengli, Lin Haofeng, Liu Meiqin, Jiang Rendi
Zhou Peng. #WIV1 #RsSHC014S
The invention relates to the technical field of coronavirus, in particular to a BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC 014S.
The SARS-associated coronavirus #RsSHC014 can effectively replicate and propagate in a corresponding cell line and a transgenic mouse by using #hACE2 as a receptor,
RsSHC014 cannot effectively replicate and proliferate in a wild BALB/c mouse, a #hACE2 transgenic mouse is expensive and not easy to obtain, the transgenic mouse infected with the #RsSHC014 can cause brain infection and death...
...constructing a #ChimericVirus with #RsSHC014 #Spike gene on #WIV1 as a framework, replacing the Spike gene of WIV1 with the RsSHC014 Spike gene, obtaining an infectious clone plasmid of WIV1-RsSHC014S by utilizing a #BAC system, and constructing a chimeric virus genome..
mice #lungs were mixed with 100ul of ground supernatants for infection 2 times, #passaged to passage 19...The mice have obvious pathological characteristics after being infected with 19th generation virus...
the invention obtains the BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC014S..the mouse adapted strain can be well replicated & proliferated in lung tissues of wild BALL/c mouse, the lung of the mouse is changed pathologically, the mouse is dead.
Constructing a #ChimericVirus with #RsSHC014 Spike gene on #WIV1 as a framework, replacing the Spike gene of WIV1 with the RsSHC014 Spike gene by using a #ReverseGenetics technology, obtaining an infectious clone of WIV1-RsSHC014S by using a #BAC system..
Removing the BglI enzyme cutting site in WIV1 #skeleton ...
The divided E fragment of #WIV1 #backbone was amplified and BsaI #CleavageSites were introduced..
The divided E fragment of #WIV1 #backbone was amplified and BsaI #CleavageSites were introduced..
...infected mice were kept on a constant rearing basis, and were weighed daily and observed for mental status. Dissecting after infection for 3 days, and grinding all lung tissues of the mice by using a grinding tube containing a DMEM culture medium;
the mouse adapted strain can be replicated & proliferated in lung tissues of wild BABL/c mice,so that the lung of the mice generates obvious pathological changes,the mice die,& the development process of respiratory diseases caused by SARS-related coronavirus can be simulated.
CN114107232B Patent for a
BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S. Granted 19/4/22... #WIV #ZhengliShi
#WIV1 #RsSHC014S #tggtcg
#tctggg
patents.google.com/patent/CN11410…
BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S. Granted 19/4/22... #WIV #ZhengliShi
#WIV1 #RsSHC014S #tggtcg
#tctggg
patents.google.com/patent/CN11410…
👉2012 A China Patent for "A Seamless Gene Cloning Method" is granted & assigned to the
"Institute of Animal Science and Veterinary of #Hubei Academy of Agricultural Sciences."
"Institute of Animal Science and Veterinary of #Hubei Academy of Agricultural Sciences."
Summary of the invention: The technical problem to be solved by this invention is to provide a kind of fast, easy, economical and efficient #seamless gene #cloning method that does not rely on #enzyme-cut chain reaction. #Cloning #BglII
Dec 2020: @janeqiuchina interview with #Shi ..Shi’s main contribution was to provide #Baric with SHC014’s yet to be published #genomicsequence. It turned out that both the synthesized #SHC014 and the #SARS-CoV-1-SHC014 #chimera were able to infect human cells & make mice sick.
CN114107232B Patent
BALB/c mouse adapted strain of SARS related coronavirus #WIV1-RsSHC014S and application.
Why the Rush? Why only 🇨🇳? #ZhengliShi
BALB/c mouse adapted strain of SARS related coronavirus #WIV1-RsSHC014S and application.
Why the Rush? Why only 🇨🇳? #ZhengliShi
How 'fortuitous' was it for #Baric that #TCTGGG was seen to be present in #SarCoV2 ? #Pradhan #SHC014
SARS-related bat coronavirus #RsSHC014 #PREDICT #EHA -156-11-YN-SHC014 RNA-dependent RNA polymerase #mRNA . #Karesh #Lipkin #Daszak #Wolfe #Ecohealth
👉 #TCTGGG Not Present??
👉 #TCTGGG Not Present??
#TCTGGG is present in #SarsCoV2 samples, #RsSHC014 , 👉 but NOT in #RsSHCO14 PREDICT-EHA, present in #MA15 &
#MA15-SHC014 Spike, but NOT in #RaTG13 , #BtCoV/4991, nor #Banal20/52Laos.
#MA15-SHC014 Spike, but NOT in #RaTG13 , #BtCoV/4991, nor #Banal20/52Laos.
.. while the #Shi patent BALB/c mouse adapted strain of SARS related coronavirus #WIV1-RsSHC014S also shows #TCTGGG is present.
Very #uncanny that #Fauci et al acted so quickly to dismiss the #Pradhan paper 'BEFORE' it had been peer-reviewed. #TCTGGG
#TCTGGG / #UCUGGG is 'NOT' present in #BANAL-20-52/Laos/2020, but 'uncannily' the
#TGGTCGC / #UGGUCGC is!
#TGGTCGC / #UGGUCGC is!
https://twitter.com/jhas5/status/1477482687313108993?t=UvMkIZynenT6MQ7q--DNOw&s=19
👉So #TGGTCGC / #UGGUCGC is present in #Banal-20-50/Laos/2020, AND also in #Sars #Rs4231 , #Rs4237 , #Rs4255 & #Rs3367 . #InstitutPasteur #Eloit #Daszak #Wang & #Shi . #Uncanny
#Shi #WIV #RsSHC014 #Baric 👉 #PREDICT #EcoHealth #Daszak #Lipkin #Karesh
👉#Wolfe #Metabiota #Biden #Pradhan #TCTGGG #Uncanny
👉#Wolfe #Metabiota #Biden #Pradhan #TCTGGG #Uncanny
#TCTGGG #TGGTCGC #RaTG13 #Pradhan #Uncanny
https://twitter.com/stevenemassey/status/1464284407108358146?t=Oyrn0Q7CrCBqeKb4UQtgMw&s=19
The #RaTG13 Complete Genome Version 1 was first uploaded Jan 29 2020. The Sequence table showed that #TCTGGG is present twice. It also shows that #TGGTCGC is 'NOT' present! #Uncanny
On Oct 13 2020, Version 2 of the #RaTG13 Complete Genome is uploaded and the Sequence table shows that the 2 previous references to #TCTGGG are now 'NOT' present. While #TGGTCGC is now 'PRESENT'. #Uncanny #Pradhan
#Baric #Yount #NoSeem METHODS & COMPOSITIONS FOR INFECTIOUS CDNA OF SARS CORONAVIRUS... #Pradhan #TCTGGG #HIV #Fauci #Uncanny 🤫
Why ignore a 'Banned' non 'peer-reviewed paper? #Pradhan #TCTGGG #QTNSSPRRA #Fauci @TheLancet
https://twitter.com/tommy_cleary/status/1533744869218619394?t=fyTFjzl6zsyy0sj-fND-_Q&s=19
👉 January 31st 2020:
#Uncanny similarity of unique inserts in the #2019-nCoV spike protein to #HIV-1 gp120
and Gag. #PrashantPradhan #SarsCoV2 #HIV @TheLancet #Fauci #Holmes #Baric #Shi
#Uncanny similarity of unique inserts in the #2019-nCoV spike protein to #HIV-1 gp120
and Gag. #PrashantPradhan #SarsCoV2 #HIV @TheLancet #Fauci #Holmes #Baric #Shi
On examination of the sequence alignment we found that the #2019nCoV spike glycoprotein contains 👉 4 insertions....We found that these 4 insertions are 👉unique to 2019-nCoV & are 👉 'NOT' present in any other coronaviruses analysed. #Uncanny #Pradhan #HIV
The insert 1 & insert 2 in the spike glycoprotein of #2019nCoV are 👉 100% identical to the residues mapped to #HIV-1 gp120. The insert 3 in 2019nCoV maps to HIV-1 #gp120 with gaps. The insert 4 maps to HIV-1 #Gag with gaps. #Pradhan #Uncanny
Although,the 4 inserts represent short stretches of amino acids in spike glycoprotein of #2019nCoV , the fact that all three of them share amino acid identity or similarity with #HIV-1 #gp120 & HIV-1 #Gag suggests that this is 👉'NOT' a random fortuitous finding....
..one may expect a fortuitous match for a stretch of
6-12 contiguous amino acid residues in an unrelated protein. However, it is 👉 'UNLIKELY' that all 4 inserts in the #2019nCoV spike glycoprotein fortuitously match with 2 key structural proteins of an unrelated virus #HIV-1.
6-12 contiguous amino acid residues in an unrelated protein. However, it is 👉 'UNLIKELY' that all 4 inserts in the #2019nCoV spike glycoprotein fortuitously match with 2 key structural proteins of an unrelated virus #HIV-1.
Insert 1 = #TNGTKR Insert 2 = #HKNNKS Insert 3 = #GDSSSG
Insert 4 = #QTNSPRRA #Pradhan #Uncanny #SarsCoV2 #RaTG13 #Banal20
Insert 4 = #QTNSPRRA #Pradhan #Uncanny #SarsCoV2 #RaTG13 #Banal20
13th January 2020, NC_045512.1
Sars2 isolate #Wuhan-Hu-1, complete genome.. #TNGTKR #HKNNKS #GDSSS #QTNSPRRA #Pradhan #HIV #Uncanny
Sars2 isolate #Wuhan-Hu-1, complete genome.. #TNGTKR #HKNNKS #GDSSS #QTNSPRRA #Pradhan #HIV #Uncanny
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