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| 06:56 AM | ||
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| 06:56 AM | ||
Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] | |
| Anonymous Coward User ID: 40421159  Canada03/13/2020 07:42 PM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] Since China wants to play the blame game against the US over a virus they created I updated this with the 2007 Wuhan study that confirms China created a Bat Corona psuedovirus using HIV inserts to make it bind to ACE2 receptors to human transmission. On first post. Quoting: YouAreDreaming Here is a 2008 study from the lab in Wuhan that details step by step how they took a natural SARS-bat coronavirus and modified it into a psuedovirus using HIV inserts so it can bind to ACE2 receptors. [link to jvi.asm.org (secure)] State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China1; Key Laboratory ofCell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China2;CSIRO Livestock Industries, Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center forEmerging Infectious Diseases, Geelong, Australia3; and School of Life Science, East China Normal University, Shanghai, China In this study, a human immunodeficiency virus (HIV)-basedpseudovirus system was employed to address these issues. Ourresults indicated that the SL-CoV S protein is unable to useACE2 proteins of different species for cell entry and thatSARS-CoV S protein also failed to bind the ACE2 molecule ofthe horseshoe bat,Rhinolophus pearsonii. However, when theRBD of SL-CoV S was replaced with that from the SARS-CoVS, the hybrid S protein was able to use the huACE2 for cellentry, implying that the SL-CoV S proteins are structurally andfunctionally very similar to the SARS-CoV S. These resultssuggest that although the SL-CoVs discovered in bats so far areunlikely to infect humans using ACE2 as a receptor, it remainsto be seen whether they are able to use other surface moleculesof certain human cell types to gain entry. More from the article on how the created the HIV psuedovirus to prove their thesis of human infectivity. Construction and purification of pseudoviruses.HIV-based pseudoviruseswere prepared as described previously (37). In brief, 12g each of pHIV-Luc(pNL4.3.Luc.RE-Luc) and the S-expressing plasmids (or empty vector con-trol) was cotransfected into 2106293T cells in 10-cm dishes. After 8 h, themedium was replaced with fresh medium. Supernatants were harvested at 48 hposttransfection, clarified from cell debris by centrifugation at 3,000g, andfiltered through a 0.45-m-pore-size filter (Millipore). Pseudoviruses in the cellsupernatant (4 ml) were purified by ultracentrifugation through a 20% sucrosecushion (4 ml) at 55,000gfor 60 min using a Ty90 rotor (Beckman). Thepelleted pseudoviruses were dissolved in 100l of phosphate-buffered saline(PBS) and stored at80°C in aliquots until they were used. To evaluate theincorporation of S proteins and HIV p24 into the pseudotyped viruses, 20lofpurified viruses was subjected to sodium dodecyl sulfate-polyacrylamide gel elec-trophoresis (SDS-PAGE) and Western blotting as detailed below. This equals... Made in China! Clear as mud |
| Anonymous Coward User ID: 69958482  United States03/13/2020 08:53 PM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] |
| Anonymous Coward User ID: 71408469 United States03/13/2020 10:05 PM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] This right here. This is warfare plain and simple. Trump needs to wake the fuck up. ~~~~ Trump knows precisely what’s going on. Trump don’t know shit about fuck. he is still the President. and you are what? |
| Anonymous Coward User ID: 54341606 United States03/13/2020 10:08 PM Report Abusive Post Report Copyright Violation | |
| Anonymous Coward User ID: 78613521 United States03/13/2020 10:17 PM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] Since China wants to play the blame game against the US over a virus they created I updated this with the 2007 Wuhan study that confirms China created a Bat Corona psuedovirus using HIV inserts to make it bind to ACE2 receptors to human transmission. On first post. Quoting: YouAreDreaming Here is a 2008 study from the lab in Wuhan that details step by step how they took a natural SARS-bat coronavirus and modified it into a psuedovirus using HIV inserts so it can bind to ACE2 receptors. [link to jvi.asm.org (secure)] State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China1; Key Laboratory ofCell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China2;CSIRO Livestock Industries, Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center forEmerging Infectious Diseases, Geelong, Australia3; and School of Life Science, East China Normal University, Shanghai, China In this study, a human immunodeficiency virus (HIV)-basedpseudovirus system was employed to address these issues. Ourresults indicated that the SL-CoV S protein is unable to useACE2 proteins of different species for cell entry and thatSARS-CoV S protein also failed to bind the ACE2 molecule ofthe horseshoe bat,Rhinolophus pearsonii. However, when theRBD of SL-CoV S was replaced with that from the SARS-CoVS, the hybrid S protein was able to use the huACE2 for cellentry, implying that the SL-CoV S proteins are structurally andfunctionally very similar to the SARS-CoV S. These resultssuggest that although the SL-CoVs discovered in bats so far areunlikely to infect humans using ACE2 as a receptor, it remainsto be seen whether they are able to use other surface moleculesof certain human cell types to gain entry. More from the article on how the created the HIV psuedovirus to prove their thesis of human infectivity. Construction and purification of pseudoviruses.HIV-based pseudoviruseswere prepared as described previously (37). In brief, 12g each of pHIV-Luc(pNL4.3.Luc.RE-Luc) and the S-expressing plasmids (or empty vector con-trol) was cotransfected into 2106293T cells in 10-cm dishes. After 8 h, themedium was replaced with fresh medium. Supernatants were harvested at 48 hposttransfection, clarified from cell debris by centrifugation at 3,000g, andfiltered through a 0.45-m-pore-size filter (Millipore). Pseudoviruses in the cellsupernatant (4 ml) were purified by ultracentrifugation through a 20% sucrosecushion (4 ml) at 55,000gfor 60 min using a Ty90 rotor (Beckman). Thepelleted pseudoviruses were dissolved in 100l of phosphate-buffered saline(PBS) and stored at80°C in aliquots until they were used. To evaluate theincorporation of S proteins and HIV p24 into the pseudotyped viruses, 20lofpurified viruses was subjected to sodium dodecyl sulfate-polyacrylamide gel elec-trophoresis (SDS-PAGE) and Western blotting as detailed below. This equals... Made in China! BRILLIANT catch!!! Excellent work. That proves the Chinese were conducting all sorts of dubious experiments, including trying to combine HIV and SARS type viruses.  |
| Anonymous Coward User ID: 78599460  Ireland03/13/2020 10:26 PM Report Abusive Post Report Copyright Violation | |
YouAreDreaming
(OP) User ID: 78123874 Canada03/14/2020 11:45 AM Report Abusive Post Report Copyright Violation | |
| serena28
User ID: 78507281 United States03/15/2020 12:25 AM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] (December 19, 2017) The US government is lifting a ban on engineering deadly viruses to make them more dangerous Quoting: Anonymous Coward 77201416 [link to www.sciencemag.org (secure)] [link to www.businessinsider.com (secure)] [link to grants.nih.gov (secure)] DAMN! Gain-of-function. They/us have purposefully been trying to make dangerous viruses MORE communicable. Couldn't see why that might be a problem... |
| YouAreDreaming
(OP) User ID: 78123874 Canada03/15/2020 07:54 AM Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] (December 19, 2017) The US government is lifting a ban on engineering deadly viruses to make them more dangerous Quoting: Anonymous Coward 77201416 [link to www.sciencemag.org (secure)] [link to www.businessinsider.com (secure)] [link to grants.nih.gov (secure)] DAMN! Gain-of-function. They/us have purposefully been trying to make dangerous viruses MORE communicable. Couldn't see why that might be a problem... In know right... fucking Chinese creating weapons of mass destruction in the form of bioengineered viruses and the world coughs and says it's just the flu. Utter bullshit! The deception and lies are enormous! |
Astroshill
Senior Forum Moderator 03/16/2020 03:28 PM  Report Abusive Post Report Copyright Violation | Re: Time to wake up GLP... nothing burgers, shills, idiots... we have proof! Tick tock... tick tock...[Updated] A finger-print protein of a Bat Sars coronavirus was uploaded to NCBI in 2018 by the Institute of Military Medicine Nanjing on January 5th, 2018 that 100% matches today's current Covid-19 envelope protein. Quoting: YouAreDreaming In 2 years, the chances of this protein not undergoing mutations and matching this 2018 virus is next to statistically impossible. In several current submissios from other countries in 3 months, there have been several mutations of this envelope protien. Although Astro points out the entire Genome of 2018 is different from the 2019 December strain but are 89% similar that suggests not the exact same species. This also further indicates that the 2018 protien could have been copied over to the current strain which wouldn't happen in nature but definately happen in a lab as per Dr. Gary Ades. I'm not happy you decided to further edit the OP to try to minimize the effort I put into properly researching this. Just because the amino acid sequence is identical does NOT mean there were no mutations and that the genetic sequence is still identical. I know you guys aren't molecular biologists, but amino acids are coded for by a set of three nucleotides called a codon. Multiple codons can code for the same amino acid. [link to www.ncbi.nlm.nih.gov (secure)] This means that single mutations that change a single nucleotide of the virus genome do not necessarily change the amino acid that codon will code for. These are called synonymous mutations. There ARE synonymous mutations present in the 2019-nCoV virus within the region that codes for the envelope protein (AVP78033.1). This protein is highly conserved between multiple strains of the virus, this is expected when the protein being encoded is vital to function and most mutations result in dysfunction. Many other strains have >90% similarity in genetic sequence of this protein. In the case of 2019-nCoV, the similarity is 98.68%. Here is the result of a BLAST on the genetic sequence showing the alignment between the older virus' envelope sequence and the current virus: [link to drive.google.com (secure)] The spots where there is not a vertical line is where the mutations are located.  |
| Kilmarnock User ID: 43889506 United States03/17/2020 07:55 AM Report Abusive Post Report Copyright Violation | |
| Anonymous Coward User ID: 72878711 United States10/27/2020 01:19 PM Report Abusive Post Report Copyright Violation | |
