🛑🛑🛑➡️ The results of our 5-year SARSr-CoV surveillance in a cave inhabited by multiple species of horseshoe bats in Yunnan.
👉The Study reveal that the SARSr-CoVs circulating in this single location are highly diverse in the S, ORF3 and ORF8 gene!👇

1)A large number of
2) SARS-related coronaviruses (SARSr-CoV) have been detected in horseshoe bats since 2005 in different areas of China. However, these bat SARSr-CoVs show sequence differences to SARS coronavirus (SARS-CoV) in different genes (S, ORF8, ORF3, etc.). Below we report the results
3) of our 5-year SARSr-CoV surveillance in a cave inhabited by multiple species of horseshoe bats in Yunnan Province, China. The full-length genomes of 11 recently discovered SARSr-CoV strains, along with our previous findings, reveal that the SARSr-CoVs circulating in this
4) single location are highly diverse in the S, ORF3 and ORF8 gene.
Importantly, strains with high genetic similarity to SARS-CoV in the N-terminal hypervariable domain (NTD) and receptor binding domain (RBD) of the S1 gene, respectively in the ORF3 and 👉ORF8 region, were all
5) discovered in this cave. Furthermore, we report the first discovery of bat SARSr-CoV highly similar to human SARS-CoV in ORF3b and in the👉 ORF8a and 8b division. Furthermore, the SARSr-CoV strains from this cave were more closely related to SARS-CoV in the non-structural
6) protein genes ORF1a. Recombination analysis shows evidence of frequent recombination events within the 👉S gene and around the 👉ORF8 among them SARSr-CoV.
We hypothesize that the direct progenitor of SARS-CoV may have originated after 👉sequential recombination events
7) between the precursors of these SARSr-CoV.
Cell entry studies have shown that three recently identified SARSr-CoVs with different S protein sequences are all capable of using human ACE2 as a receptor, further demonstrating the close relationship between the strains in this
8) cave and SARS -CoV. This work provides new insight into the origin and evolution of SARS-CoV.👇
ncbi.nlm.nih.gov/pmc/articles/P…
9)Amino acid sequence comparison of the S1 subunit (corresponding to aa 1–660 of the spike protein of SARS-CoV).👇

The receptor-binding domain (aa 318–510) of SARS-CoV and the homologous region of bat SARSr-CoVs are indicated by the red box. The key aa residues involved in
10) the interaction with human ACE2 are numbered on top of the aligned sequences. SARS-CoV GZ02, BJ01 and Tor2 were isolated from patients in the early, middle and late phase, respectively, of the SARS outbreak in 2003. SARS-CoV SZ3 was identified from civets in 2003.
11) SARSr-CoV Rs 672 and YN2013 were identified from R. sinicus collected in Guizhou and Yunnan Province, respectively. SARSr-CoV Rf1 and JL2012 were identified from R. ferrumequinum collected in Hubei and Jilin Province, respectively. WIV1, WIV16, RsSHC014, Rs4081, Rs4084,
12) Rs4231, Rs4237, Rs4247, Rs7327 and Rs4874 were identified from R.sinicus, and Rf4092 from R. ferrumequinum in the cave surveyed in this study.
13) Alignment of nucleotide sequences of ORF8 or ORF8a/8b.👇
The start codons and stop codons of ORF8, 8a and 8b are marked with black boxes and the forward and reverse arrows, respectively. The deletion responsible for the split ORF8a and 8b in human SARS-CoV BJ01, Tor2 and bat
14) SARSr-CoV Rs4084 is marked with red boxes. See the legend for Fig 3 for the origin of various sequences used in this alignment.
15) Functional characterization of diverse ORF8 and ORF8a proteins of bat SARSr-CoVs.👇

(A) The ORF8 proteins of SARS-CoV and bat SARSr-CoVs induces the ATF6-dependent transcriptional activity. HeLa cells were transiently transfected with the pcAGGS expression plasmids
16)of the ORF8 of SARS-CoV GZ02, bat SARSr-CoV Rf1, WIV1 and Rf4092 and the reporter plasmid 5×ATF6-GL3 for 40h. Control cells were co-transfected with the reporter plasmid and the empty pCAGGS vector for 24h, and treated with or without TM (2μg/ml) for an additional 16h.
17) The cell lysates were harvested for dual luciferase assay and data are shown as the average values from triplicate wells. (B) The ORF8a proteins of SARS-CoV and bat SARSr-CoV triggered apoptosis. 293T cells were transfected with the expression plasmids of the ORF8a
18) of SARS-CoV Tor2 and bat SARSr-CoV Rs4084 and a pcAGGS vector control for 24h. Apoptosis was analyzed by flow cytometry after annexin V staining and the percentage of apoptotic cells were calculated. Data are shown as the average values from
19) triplicate cells. Error bars indicate SDs. * P<0.05. 👇
🛑🛑🛑➡️ This passage is more important👇
➡️Recombination analysis shows evidence of frequent recombination events within the S gene and around the ORF8 among them SARSr-CoV.
We hypothesize that the direct progenitor of SARS-CoV may have originated after sequential recombination events between the precursors of these
SARSr-CoV.
Cell entry studies have shown that three recently identified SARSr-CoVs with different S protein sequences are all capable of using human ACE2 as a receptor, further demonstrating the close relationship between the strains in this cave and SARS -CoV.
SARS COV 2 ORF8, and the ricombination genome with utilized of the Ferrets (intermediate animal).👇

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🛑🛑🛑➡️1) This work was supported, in part, by a fund-raising effort in which approximately 330 persons contributed funds in support
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2) CC BY-NC-SA. Any derivative use of this dossier must be made public for the benefit of others. All documents, references and
disclosures contained herein are subject to an AS-IS representation. The author does not bear responsibility for errors in
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