That's why healthcare was wrecked and the abomination of ObamaScare was installed. Obama's own man called it "the stupidity of the American voter" - and the idiots STILL believe in Obama. SMH
Posted on 07/15/2023 7:48:33 PM PDT by SeekAndFind
And they KILLED THOUSANDS of OLD PEOPLE in the process of getting rid of Trump....it was a TWO-FER!
That's why healthcare was wrecked and the abomination of ObamaScare was installed. Obama's own man called it "the stupidity of the American voter" - and the idiots STILL believe in Obama. SMH
Daszak
Fauci killed millions of people and ruined the lives of many millions more. Despite this, he is still not only free, but he is living the lavish life of an aristocrat.
Compare this to the J6 prisoners in solitary for wrongthink.
Yep. Nobody in DC will drill down on this....because they all know they will find that it was conveniently leaked “on purpose by accident”
“There is no evidence that the SARS-CoV-2 virus is anything other than a natural bat virus that has since adapted to infecting humans. If it had been deliberately altered, the evidence of that would be very clear in its RNA, because nucleic acids cannot be altered without leaving a trail of evidence.”
Wow, are you familiar with Golden Gate Cloning? Do you know what a restriction enzyme is? have you heard of a restriction enzyme recognition sequence? Do you realize that there were several infectious clones of bat coronaviruses created before the pandemic, and the method used to create them was published?
Using well known cloning techniques, viruses can be cut into pieces and re-assembled.
Here are my links in support.
https://www.biorxiv.org/content/10.1101/2022.10.18.512756v1
https://www.neb.com/tools-and-resources/selection-charts/type-iis-restriction-enzymes
https://www.neb.com/tools-and-resources/feature-articles/everything-you-ever-wanted-to-know-about-type-ii-restriction-enzymes
https://www.nature.com/scitable/topicpage/restriction-enzymes-545/
https://www.addgene.org/protocols/subcloning/
https://www.labxchange.org/library/pathway/lx-pathway:27bdb1d1-b7cf-4d7d-95fb-e28e9c6e21f9/items/lx-pb:27bdb1d1-b7cf-4d7d-95fb-e28e9c6e21f9:html:2606f1c1
https://parts.igem.org/Help:Assembly/Scars
Type IIS cleavage domains have no inherent sequence-specificity, and so the sequence of the overhang they generate varies from one recognition site to another.
Fragments produced by Type IIS-digestion of natural DNA molecules generally have different overhangs, therefore, and will not anneal to one another.
However, if the sequence of the overhang is predetermined, by designing it into a PCR primer, for example, then it can be made to complement another and to be directional.
This feature is used to great advantage in ‘Golden Gate’ assembly where multiple fragments can be stitched together in the correct order and orientation in a single ligation.
The Type IIS enzymes, BsaI (GGTCTC 1/5), and BsmBI (CGTCTC 1/5), are very popular for this application.
The advantage of using Type IIS enzymes for assembly is that the recognition sequence can be placed in the primer on either side of cleavage site.
If placed ‘inside’, 3’ to the cleaved end, it will be retained in the construct and can be re-used subsequently.
If placed outside, 5’ to the cleaved end, it will be lost, leading to a ‘scar-less’ assembly.
https://blog.addgene.org/plasmids-101-golden-gate-cloning
As per the below, the advantage of scarless cloning is that if 2 pieces of the plasmid are cut, they may re-connect in their original orientation. If there is a “scar”
(a difference in the re-connected parts from their original sequence) the restriction enzyme cannot cut them a second (or more) time. But if the re-connected
parts do not have a “scar” the restriction enzyme will cut the plasmid again in the same way, increasing the chances of the parts remaining separated in the desired way (?)
Advantages of Golden Gate cloning
Golden Gate cloning is one of the easiest cloning methods in terms of hands-on time, as digestion and ligation can be done in one 30-minute reaction.
The destination vector and entry vector(s) are placed in a single tube containing the Type IIS enzyme and ligase.
(below - advantage of not leaving a scar)
******Although the original destination vector + insert may spontaneously religate, this transient construct retains functional Type IIS sites and will be re-digested.***
In contrast, formation of the desired ligation
product is irreversible because this construct does not retain the enzyme recognition sites. As a result, the ligation process is close to 100% efficient. Another
strength of Golden Gate cloning is its scalability. Unique 4 base overhangs can be used to assemble multiple fragments - up to 10 fragments are commonly
assembled in a single reaction! These overhangs specify the desired order of fragments, and the loss of enzyme recognition sites after ligation favors formation
of the construct of interest. Although efficiency may decrease with an increased number of fragments, or the ligation of very small/very large fragments, these
problems can be overcome by screening a higher number of potential clones. Golden Gate assembly has a few advantages over other cloning methods.
Exonuclease-based methods like Gibson assembly require 20-40 bp of homology at the ends of DNA fragments to specify assembly order, so fragments
with 5’ or 3’ sequence homology cannot be assembled using this method, but can be assembled with Golden Gate. The popular Gateway cloning system
produces constructs with an attB recombination scar encoding eight amino acids, but Golden Gate assembly can be designed to be scarless. Golden Gate
assembly is also less expensive than many commercial cloning methods.
https://www.neb.com/applications/cloning-and-synthetic-biology/seamless-cloning
Another advantage of Type IIs - the overhang can be 6nt with a 6nt recognition sequence which is the idea length - 4^6 is 4096 which is about 1/7th of the virus length.
with 2^5 there are like 28 sites in the virus, with 2^7 there is likely only 1. Regardless of the length of the overhang, there is a big chance of matching the wrong overhang
if there are multiple fragments created with the same type II, because all the type II sites (likely 7 of them with a 6nt type II) match each other.
With type IIs you can have a 6nt recognition sequence and a 6nt overhang (?). With type II if you have multiple fragments created at once all the overhangs are the same so
the fragments will likely not assemble in the right order. With type IIS the overhangs are different so much more likley to assemble in the correct order. You would have to
find multiple type II to make 6 fragments, and each one would have to not interfere with the others, and only affect one site, which is unlikley since there are usually
7 sites for any 6nt recognition sequence. 6nt with type II produces inverted fragments and loops. 7nt not enough sites (only 1 usually), and why would they
be anywhere near the desired location? plus you would need 6 different ones, How many 7nt enzymes exist?
Scobey creation of MERs clone
https://pubmed.ncbi.nlm.nih.gov/24043791/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3791741/
https://www.thermofisher.com/us/en/home/brands/thermo-scientific/molecular-biology/thermo-scientific-restriction-modifying-enzymes/restriction-enzymes-thermo-scientific/fastdigest-thermo-scientific/type-iis-restriction-enzymes.html
Cloning with Type IIS restriction enzymes
- if multiple fragments have idential ends, then number of erroneous fragment recominations possible grows exponentially with the number of fragments. There may be
no practical way to isolate the small fraction of recombined fragments that have recombined successfully. gel purification is based on fragment size, all the various combinations
of N fragments would have the same size but random fragment order, mixed in with recombinations of N-1 fragments, N-2 fragments, etc.Any of the fragments might form a loop or
be inverted. so 4 fragments could be ordered in 4*3*2 = 24 ways, with 2^4 = 16 possible inversion combinations.
Traditional cloning workflows involve multiple hands-on steps. Typically, a recipient plasmid vector is digested with one or more restriction enzymes,
the terminal ends may then be dephosphorylated, and finally the linearized plasmid is gel purified. In parallel, the fragment to be cloned is also digested and gel
purified, or alternatively, PCR amplified, digested to create compatible ends, and column purified. The two molecules are then treated with DNA ligase and transformed
nto E.coli.**** Traditional restriction enzyme cloning is usually limited to inserting a single DNA fragment into a recipient vector***
In contrast, Golden Gate cloning utilizes Type IIS restriction enzymes, in combination with DNA ligase, in a single reaction tube to drive the insertion of a DNA fragment –
or several DNA fragments – into a recipient vector. Typically the reaction, performed in a thermocycler, is cycled repeatedly between the temperatures optimal for the
restriction endonuclease (37 °C) and the DNA ligase (23 °C). As a result, Golden Gate DNA assembly reduces (or eliminates) multiple hands-on steps and does not
require agarose gel purification [3].
effect of level of firal exposure on diease severity https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4342672/
- Why inverted and random fragments make it impossible to analyse virulence of virus - let’s say 1 percent of virus copies are correctly aligned and the rest have some
inversions or mis-aligned fragments or extra fragments. If this mixed bag of viruses is tested for virulence, it is impossible to know whether any disease results from the
small portion of correctly aligned viruses, or whether some of the mis-alignments are irrelevant to the potential to cause disease. The only way to know this is to purify and isolate
the correctly aligned viruses. Even if this is possible, it is a huge disadvantage to have to attempt this.
article discussing furin cleavage site and CGGCGG arginine coding
https://www.caltech.edu/about/news/the-debate-over-origins-of-sars-cov-2
https://www.pnas.org/doi/10.1073/pnas.2211107119 argugment about spike
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9173817/ claim that spike was engineered
https://www.nature.com/articles/d41586-021-01529-3 what scientsts do and do not know
- Arginine and FCS argument is simply that the FCS and arginine codons don’t exist in other COVs. That doesn’t mean they came from a lab or that they are consistent with
practices used in a lab, unless there is some reason why, in a lab you have to create a FCS with arginine coded as GCCGCC. With so many viruses in circulation
known and unknown I don’t see how a probabilty can be assigned. But Washburne is saying the recog. sites for type IIS enzymes have a pattern which is required for lab
and no other COVs have this pattern. So the wild theory can be shown false because all lab created COVs have the same pattern and none of the wild COVs have the pattern.
Washburne is saying there is an unusual pattern or feature, and that this feature is consistent with lab origin, as well as inconsistent with natural origin. The arginine argument just
says the pattern is inconsistent with wild origin, not that it is consistent with lab origin, because the GCCGCC can come from anywhere with an unkown likelihood. You would
have to look at how new codons get into new viruses, and this seems like a messy and imprecise line of argument. If GCCGCC was easier to insert in a lab, or the only possible
codon a lab could insert for some practical reason, and if there was a history of scientists inserting it, the argument would be as strong as Washburne. But we don’t see
that argument being made, just the argument that GCCGCC is weird and other COVs don’t have it.
exDemMom you write
“ The Wuhan scientists replaced the spike protein on a human coronavirus with a spike protein from a bat virus to see if the human virus could still infect human cells with a bat virus spike protein”
This simplistic description of what “Wuhan scientists” did is obviously not correct, because there are many published papers describing a variety of cloning attempts and manipulation of different viruses. Also, the spike protein itself was probably manipulated because it contained a furin cleavage site. I think SARS 1 had a furin cleavage site on its spike protein.
“Replacing a protein in an organism with an analogous protein from a different organism is not “gain-of-function,” no matter how many times conspiracy theorists claim that it is”
Not even a nice try! It depends what the protein does, correct?
And then you start talking about “vectors”. Thanks for giving a portion of the Covid 19 genome. I have the whole thing, all 32k, along with the genomes for 70 other bat coronaviruses, because the Washburne paper has a link to this data on github - see below
https://github.com/reptalex/SARS2_Reverse_Genetics
“To look for traces of vector in the SARS-CoV-2 virus,”
the phrase “traces of vector” is a clue that your post was written by someone who doesn’t speak English.
“telling you that it found no trace of vector contamination in the sequence.”
Wow tin-foil hat stuff. Nice try though, I’m sure someone will be impressed with your vector language and believe your post, but it is made-up gibberish.
continuing to spread every conspiracy theory about ... does not help the conservative cause at all. On the contrary, it has helped to promote an image of conservatives as ...
/\
I’m seeing this comment a lot about a lot of subjects lately..
Bot-ish...
I’m gonna add it to my diversion list
“... I believe..,, “
“ ...out of context..”
“ I don’t agree with your characterisation of...”
” continuing to spread every conspiracy theory about ... does not help the conservative cause at all. On the contrary, it has helped to promote an image of conservatives as ...”
What was the cdc doing with a bio weapons lab in communist china and Ukraine. Put a bag over Fauci and others at the CDC and release blow flies inside like he did with beagle dogs and ask him for the truth. Once he tells us then leave the bags attached to them all and let the insect larva finish the job.
I worked in a Help Desk job and was supporting internal employees for a large company (near 200,000 people worldwide). One day in Sept 2019 a person had a Dell battery start to expand... It bent the laptop out of shape. The battery, keyboard, mouse trackpad would need to be replaced. I had handled issues like this before as my co-workers did. Orders were placed but nearly 2 months went by before they got the repair parts and Dell showed up to fix the pc. The parts came from china.
One day earlier another co-worker had the same issue and it was completed within a few days like normal. I bet Dell ran out of parts and the delay for the later call was due to getting them from China.
I believe this wuhan virus started much sooner then we have been told.
Bio-Weapon.
I realized that early on.
[Bio-Weapon.]
I definitely think so.
For War?
For use on civilians for Population Reduction?
Perhaps both?
Yes.
https://freerepublic.com/focus/news/4168208/posts?page=9#9
A long read.
Important is that the Wuhan lab was changed from civilian control to Chinese Military purposely
Plus lots of info showing how it was purposely created and used as a Weapon of War.
Part of China’s long term plan to control the whole world.
Thanks Deep State.
[Important is that the Wuhan lab was changed from civilian control to Chinese Military purposely]
And there it is.
Why am I not surprised?
Plus all Chinese MUST do whatever #CCP dictates.
Even those here in the USA.
They have jails here in USA.
They will snatch China ppl off the street and ‘Disappear’ them.
Unless they can reprogram them.
Even if they are US citizens❗️
Just wait until they implement Social Credit Scoring
I’m thinking that’s no longer just TV stuff
It’s leading to that WARP SPEED IMO!
It’s all coming together as planned.
Check out Lance Wallneu, his weekly show on Daystar is packed with the best plan out there.
He’s got a lot of highly aware Christians he is working with.
Chuck Missler was one of them before he died.
Check this out.
https://lancewallnau.com/tag/president-trump/
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