Message from PLA Vaccine Patent.

Zhiyan-Le, 2021-03-15.
https://sites.google.com/site/zhiyanleback/2021-1/z20210315-patent-message-en

Patents of nCov S-Gene by PRC Wuhan Institute of Virology (WIV), and that of nCov vaccine by PRC PLA doctor Chen Wei team, sent a clear message that the SARS-2/Covid-19 virus can be man-made. Dr. Chen Wei especially emphasized that the single gene GC quantity and distribution have a significant contribution to the stability of the virus and its affinity with human ACE2. Such message suggests that the SARS-2 origin comes from artificial chimera WIV1 (Wuhan Institute of Virology, No. 1 virus), that is, the source of the pandemic does not come from natural animals such as bats in PRC Yunnan province or in South-East Asia communities.

First, sampling for the S-gene sequence, chosen from NIH GenBank and WIPO:

• NC-045512: Base sample, from Wuhan patient, collected at a local hospital in December 2019.
   

• WIV1: A synthetic nCov by Wuhan Institute of Virology (WIV), completed in 2015.

• RaTG13/S: Bat sample, said that was collected by WIV on September 2013 and released on February 2020, reported as the source of this pandemic.
   

•  WIV-CN101100680: S-Gene patent by WIV, 2007.

•  WIV-CN102690336: S-Gene patent by WIV, 2012.

•  BJ_WO2006136084: S-Gene patent by PRC Academy of Medical Science, 2006.

•  CW_CN111218459/DNA: Chen Wei vaccine gene sequence patent, on the production line on February 2020.

•  CW_CN111218459/S: Chen Wei vaccine virus gene patent, on the production line on February 2020.
   

•  MG916902.1: Yunnan bat sample, collected in 2012.

•  NC_030886.1: Yunnan bat sample, collected in 2014.
   

The above samples, and their single gene (basic gene) GC and AT of the S-protein, can be found in NIH GenBank and WIPO, as follows:

PS-Fig.01:


It can be seen that the frequency of single genes in the two samples of WIV1 and bat RaTG13 are the same, and they are the closest to the Wuhan patient sample. The natural bat samples are not only significantly different from patient sample but also from each other.


Removing patents that obviously man-made, and using the single gene quantity ratio of the total number (BP) and using the gap of mutations, we have the following distribution result:

PS-Fig.02:


In the above figure, the upper table contains ratios of single gene ACGT frequency to the total number of gene sequence length (BP). Indeed, GC and C+G have more than 60% of the total, just like what PLA Doctor Chen Wei said, their quantity is important in terms stability and affinity of the SARS-2 virus.
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The lower table and chart demo the ratio gaps or distance between bat samples and the Wuhan patient sample (basic reference sample). Clearly, the two natural bat samples (2012 and 2015) have distances too big to say they are the origin of SARS-2.

However, the other two, RaTG13 and WIV1, have a very close ratio gap with the Wuhan patient sample, thus, they should be considered as the origin of SARS-2 virus. As their ratios and gaps are the same, then, which one is or is not the origin?

Their provider, Shi Zhengli and her team of PRC-WIV, stated that nCov carried by RaTG13 does not directly jump to humans. In contrast, in March 2016, the NAS (US National Academy of Science) publication warned that WIV1 is a synthetic nCov and can directly jump to humans, causing big pandemic and economic loss, as well as changing life-styles. Now, the NAS warning has been becoming a huge reality that the whole world is facing. Thus, WIV1 must have been the real origin of SARS-2.

More over, the alignment between RaTG13 and natural bat samples has a very low identity ratio. Plus, in its updated version, RaTG13 appears having 15BP gene sequence which obviously is a part of sgRNA used for gene-editing purposes. Thus, the mystery RaTG13 is very likely a artificially synthesized object to cover up the real origin WIV1.

About ACE2

In years before this pandemic, the PRC [mainstream scientist community] had a popular research movement, called [crossing-species transmission], meaning to change those virus that do not directly jump to humans into those that can directly transmit to human. The research movement, including the above patents and WIV1, has something common: Applying ACE2 interaction to reconstruct virus genetic structure for gain of functions in terms of direct transmission. And the PRC authorities have taken those successful research projects as an example to say the China is leading the world and is surpassing America in terms of biotechnology.

Below is an overview of the expression frequency of the receptor ACE2 in different populations around the world, data are released by the US NIH GenBank on April 2020:

PS-Fig.03:


Again, we see the picture described by PRC PLA Doctor Chen Wei: Single or base genes G and C have a significant greater quantity in protein expression, and A>G and C>T allelic mutations can produce obvious changes with energy moving in opposite directions.

It is very much worth noting that, at this moment of global pandemic has continued over a year, the ACE2 expression frequency of East-Asians is obvious higher but the mutation ratio is lower than others. And the biggest portion of the East-Asians are Chinese Han nationality. This may well explain why the pandemic and virus-mutations are much stronger in Europe and Americas but much weaker in China: The fitness between SARS-2 virus and ACE2 in China had already reached a certain degree before the pandemic went to global, thus, both the virus and ACE2 can stay comparatively more stable in China than that in other geo-populations. In short, the ACE2 mutations can also tell that the SARS-2 origin is in China.


The Significance re Codon Study

Codon studies lead to the same result, that is, the SARS-2 origin is artificial chimera WIV1 rather than RaTG13 or other bats.

Significance re Codon study: So far, observation and analysis of SARS-2 similarity have mostly focused on single gene alignment or identities. However, like all living objects, the SARS-2 virus survival and functioning work with Codons, of which each contains three base genes. If one of them changed, the whole Codon operation may change. Therefore, it is necessary to conduct Codon study in addition to current approaches.

Below is the raw Codon data of the complete genetic sequence regarding the said samples.for easier graphing, I used short names, they are:

WH-01: Wuhan patient number sample.
WIV1: Same as before.
RaTG13: Same as before.
Bat-2012/Bat-12: Bat samples collected in PRC Yunnan, 2012.
Bat-2014/Bat-14: Bat samples collected in PRC Yunnan, 2014.

The following figure shows the Codon occurring frequency difference or gaps between those samples and the patient sample:

PS-Fig -4:


If the gap of a sample is zero or the nearest to zero, meanwhile, each of its Codons has the smallest gap (distribution), then that sample should be considered as the SARS-2 origin. Other wise, it should be ruled out.

Clearly, the two natural bat samples (Bat-12 and Bat-14) have big gaps for both frequency and Codon distribution, thus, they are not the SARS-2 origin.

The sample RaTG13 has a big difference frequency and bigger distribution gaps in many Codons. Plus, its provider, Dr. Shi Zhengli of PRC-WIV, said that its virus cannot directly jump to humans. Thus, this sample should be ruled, although it has 96.2% identity ratio in terms of single gene alignment.

As for the sample of artificial chimera WIV1, it has the smallest gaps for both occurring frequency and distribution. Plus, it can directly jump to humans and cause huge pandemic, which was warned by the National Academy of Science, in March 2016. Thus, this sample, WIV1 of a lab-produced new corona-virus, should be considered as the real source of SARS-2 pandemic, although its identity ratio by single-gene alignment is lower than that of RaTG13.

Further, looking into the three base genes of all Codons, the WIV1 is also has the nearest similarity to the patient sample. Please see below figure:

PS-Fig. 05:


Regarding the average of frequency and distribution of the Codon three base genes, WIV1 has a 0.99468 identity ratio and RaTG13 has 0.97972, the difference is about 1.49%. The complete genetic sequences of those samples have Codons around 10000 to 10050 or so, that is, the difference 1.49% means 10000 x 1.49% >= 120 different Codons. One Codon gap or change can impact on the entire genetic network operation, how about 120 Codon gaps or changes? Its impact would be significantly very big. Thus, the Codon study re SARS-2 origin research cannot be ignored.

Based on the above figure, here are the average frequency gaps among Codon base genes:
 

Average Frequnce and Distribution for Base Genes
Average Frequence for Each Base Gene of Codons						Distance from Patient Sample WH-01
	WH-01	WIV1	RaTG13	Bat-12	Bat-14		WIV1	RaTG13	Bat-12	Bat-14
A	3038.33	2884.67	2975.33	2342.33	2552.33	A	-153.7	-63.0	-696.0	-486.0
C	1868.00	2020.67	1836.67	1462.67	2135.67	C	152.7	-31.3	-405.3	267.7
G	1983.67	2098.00	1948.67	1936.00	2419.00	G	114.3	-35.0	-47.7	435.3
T	3267.00	3099.67	3190.33	3250.00	2946.00	T	-167.3	-76.7	-17.0	-321.0
Average Gap:							-13.5	-51.5	-291.5	-26.0

The calculation is a simple arithmetic difference, for example, [A], the average frequency value of WIV1 is 2884.67, minuses the 3038.33 of the patient number sample WH-01, the average difference is [-13.5]. In comparison, this is the smallest gap; while such gap of RaTG13 is as high as [-51.5]. Obviously, WIV1 should be considered as the SARS-2 origin, not RaTG13 or other bats.

In short, single gene alignment study is necessary and, at the same time, Codon study is also important or should be seriously put into both academic and practical purposes in coping with this SARS-2 pandemic.

Reference Materials:

Name used by this essay: WH-01.
NCBI Reference Sequence: NC_045512.1
Wuhan seafood market pneumonia virus isolate Wuhan-Hu-1, complete genome
https://www.ncbi.nlm.nih.gov/nuccore/NC_045512.1

Name used by this essay: WIV1.
GenBank: KF367457.1
Bat SARS-like coronavirus WIV1, complete genome
https://www.ncbi.nlm.nih.gov/nucleotide/KF367457.1

Name used by this essay: RaTG13.
GenBank: MN996532.1
Bat coronavirus RaTG13, complete genome
https://www.ncbi.nlm.nih.gov/nuccore/MN996532.1?report=genbank

Name used by this essay: Bat-2012/Bat-12.
GenBank: MG916902.1
Bat coronavirus BtCoV/Rh/YN2012 isolate BtCoV/Rh/YN2012_Rs4125, complete genome
https://www.ncbi.nlm.nih.gov/nuccore/MG916902.1

Name used by this essay: Bat-2014/Bat-14.
NCBI Reference Sequence: NC_030886.1
Rousettus bat coronavirus isolate GCCDC1 356, complete genome
https://www.ncbi.nlm.nih.gov/nuccore/NC_030886.1


Raw Data from The Above Reference




The Chinese version:
https://sites.google.com/site/zhiyanleback/2021-1/z20210315-patent-message 

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