A Manmade SARS-2 (e-Mail to Dr. Collins (NIH)).
by Zhiyan-Le, 2020-05-11.


Re: SARS-2 Origin and NIH Funding.
From: Zhiyan-Le.
To: Dr. Collins.
Date: Mon 5/11/2020 3:29 AM..

I have learned that NIH stopped funding for EcoHealth Alliance and its bat-coronavirus projects with PRC Wuhan Inistitute of Virology.

That is a right decision. As an American citizen, I want my tax money to benefit American people first. However, the president of EcoHealth Alliance, Dr. Daszak (and his Chinese partners), has done the opposite, since this SARS-2 outbreak.

For example, Dr. Daszak has repeatedly said to the public that there is no such thing as manmade corona-virus. PRC state-run media have been heavily quoting his message and labeling different views as conspiracy theory. As you may already know, the PRC government has censured SARS-2 origin academic research, i.e., not open to the public.

Fact is, manmade corona-virus is a popular business in China and around the world. One example:

Patent ID: CN102690336A.
Patent Title: Bat SARS-like coronavirus spike protein immunity determining area and preparation method and application thereof.
Inventor: Shi Zhengli, etc. Filed at 2012-06-11, by Wuhan Institute of Virology, PRC.
URL: https://patentimages.storage.googleapis.com/6d/52/89/cdfed23ddbb51c/CN102690336A.pdf 

The patent claim content: by applying genetic engineering tools, to make a sort of bat-SARS-like S-Gene and its proteins that can be engineered and reproduced for corona-virus usages at any time.

Following the SARS-2 outbreak, the PRC Patent Office confirmed that the patent exists and is available. Please see: http://www.cnipa.gov.cn/docs/2020-02/20200214204433302086.pdf  (Note: Most of websites in PRC contain cloud-computing mul/spy-ware.).

As we know, patent/genetic engineering and its products are manmade. Natural things cannot be patented by law.

More over, taking the patent S-Gene (4126 bp) as the root/query, compare it with SARS-2 and SARS-like corona-virus, the result is as the following:



Regarding the corona-virus S-Gene, its protein stays much more stable than that in RNA format. Thus, it would be absolutely normal that the HuB(2006) has a high identity percent and, along with time goes by, the others have lower identities with the patent genome.

The question is: going to details, it clearly appears that the other identical gaps are at the same location with the same ratio, as well as have the same saw-toothed curves. That is, a uniformed mutation among human, bats and lab corona-virus has happened just in a few years.

A natural process re mutation is random. In a short period of few years, a uniformed genetic mutation among different hosts or holders can happen only by artificial, or manmade, work. As mentioned before, the corona-virus made by Wuhan Institute of Virology can directly jump to humans, but that of natural animals such as of bat cannot.

It is worth of noting that WIV1-CoV can directly jump to humans. Also, the Patent S-Gene and WIV1 S-Gene have higher identity than that of Patent x RaTG13. That is, WIV1 is much more significant in terns of the virus transmission and spreading.

Figure 1: Uniformed Mutation and Gap between WIV1 and WIV02.


That is why I have mentioned that, if someday someone found RaTG13 were genetically modified, I would not be surprised.


As a long term co-author with Dr. Shi Zhengli of Wuhan Institute of Virology, Dr. Daszak of course knows, not speaking of that his co-writing papers have citied, the said info/data..

Therefore, re the SARS-2 is artificial or manmade, what Dr. Daszak has said is seriously misleading, or, knowingly misleading.

I have also learned that, in an interview program, Dr. Fauci responded that there is no scientific evidence to tell the SARS-2 was made in a Chinese lab. The PRC state-run media has taken his words as a strong proof to tell the world that PRC government has nothing to be accountable re the global SARS-2 pandemic.

I think it is better for him to keep silent. Reason: Dr. Fauci is a US government employee and, at the same time, he is a senior member of the Vaccine Board of the Bill Gates Foundation (a serious conflict of interest problem). Just like that in software business, i.e., computer virus and anti-virus go together, vaccine business can survive because there are virus-caused diseases, whether the virus comes from nature or lab. Such business or personal relationship has already been under doubts, not to mention that, without authorization, no NIH employee may speak under hi/her job title. Otherwise, I wish to know if NIH authorized Dr. Fauci to say so.

Thank you for your attention.


Attachment:

1]: The PRC Patent Office Confirmed that CN102690336A Exists and is Available.

抗击新型冠状病毒肺炎专利信息研报
国家知识产权局抗击新型冠状病毒肺炎专利信息分析课题组
2020年2月14日
http://www.cnipa.gov.cn/docs/2020-02/20200214204433302086.pdf 
（一）涉及的冠状病毒种类、疫苗技术分类及免疫表位
1. 涉及的冠状病毒种类： SARS 疫苗 87 项，MERS 疫苗 10 项、HCoV－HKU1/NL63/ 蝙蝠冠状病毒疫苗 5 项，冠状病毒减毒疫苗设计 1 项。结果显示：有多项专利公开了源自人群和野生动物的新型冠状病毒或新型毒株，CN102690336 中提及的蝙蝠冠状病毒与此次新型冠状病毒同源性最高，研究了其可以作为抗原的 S 蛋白/基因片段。如果能对发现的新种类冠状病毒及时分析和研究，对于疫情出现后的疫苗研发应有相当的帮助。

2. 抗体的筛选制备方法的改进
（1）抗体表位的选择上以 S、NP 蛋白为主
CN102690336A 将蝙蝠 SARS 样冠状病毒的刺突蛋白（S蛋白）切割成多段，免疫动物后，利用完整 S 蛋白的单克隆抗体鉴定小鼠抗 S 单克隆抗体表位，并制备相应的检测用抗体。CN100504391C 中利用基因工程重组抗原，获得了 SARS冠状病毒 S、N、M、E 蛋白，并制备了偶联上述蛋白的抗体的免疫微球。JP2017145246A 以 MERS 冠状病毒最为保守且明显区别于其他冠状病毒的 NP 蛋白肽作为免疫原制备检测抗体。WO2019066389A1 将 MERS 冠状病毒的 NP 蛋白的 N 端和 C端构建融合蛋白，免疫小鼠并筛选单克隆抗体，用于 MERS病毒感染的检测。上述对血清抗原的检测相对于血清抗体的检测，能够在检测对象病毒感染初期即作出诊断，在病毒暴发高峰阶段半定量地区分阳性或阴性样本。#


2]: The Patent CN102690336A.

蝙蝠sars样冠状病毒刺突蛋白免疫决定区及制备方法和用途
CN102690336A
Inventor周鹏韩正刚石正丽
2012-06-11 Application filed by 中国科学院武汉病毒研究所
https://patentimages.storage.googleapis.com/6d/52/89/cdfed23ddbb51c/CN102690336A.pdf  
Abstract：：
本发明公开了蝙蝠SARS样冠状病毒刺突蛋白免疫决定区及制备方法和用途，其步骤是：A、以蝙蝠SARS样冠状病毒S基因为模板设计引物，扩增；B、通过上述扩增后得到的片段用BamHI和XhoI酶切后连接于表达载体pET32a上，并测序确定无误；C、将重组质粒纯化后，转化BL21感受态细胞，挑取单克隆进行培养，在终浓度为0.3mMIPTG的培养基中30度诱导；收集菌体超声波破碎后进行纯化，用HisTag纯化试剂盒进行纯化，纯化后在SDS-PAGE中检测蛋白的纯度，得到目的蛋白。该方法简单易行，操作方便，易于生产；该肽段具有最好的免疫原性，在鉴定小鼠抗S单克隆抗体表位中的应用方法特异性好，操作简单，易于重复。

Description
蝙蝠SARS样冠状病毒刺突蛋白免疫决定区及制备方法和用途

技术领域
[0001] 本发明属于生物技术领域，更具体涉及ー种蝙蝠SARS样冠状病毒刺突蛋白(Rp3-S)免疫决定区，同时还涉及ー种蝙蝠SARS样冠状病毒刺突蛋白（Rp3-S)免疫决定区的制备方法，还涉及ー种蝙蝠SARS样冠状病毒刺突蛋白（Rp3-S)免疫决定区的用途。

发明内容
[0005] 本发明的目的是在于提供了ー种蝙蝠SARS样冠状病毒（Rp3)刺突蛋白（Rp3_S)免疫决定区蛋白，其氨基酸序列为SEQ ID NO. I所示。该决定区信息为首次鉴定，可用于蝙蝠SARS样冠状病毒的特异性的诊断。
[0006] 本发明的另ー个目的是在于提供了ー种蝙蝠SARS样冠状病毒刺突蛋白（Rp3_S)免疫决定区蛋白的制备方法。该方法简单易行，操作方便，易于生产。
[0007] 本发明的再ー个目的是在于提供了ー种蝙蝠SARS样冠状病毒刺突蛋白（Rp3_S)中的S280-455多肽片段在鉴定小鼠抗S单克隆抗体表位中的应用。该方法特异性好，操作简单，实验易于重复。

SEQUENCE LISTING

<110> 中国科学院武汉病毒研究所
<120> 蝙蝠SARS样冠状病毒刺突蛋白免疫决定区及制备方法和用途
<170> PatentIn version 3.1
<213> 蝙蝠SARS样冠状病毒
<400> 1
Ile Asp Cys Ala Gln Asn Pro Leu Ala Glu Leu Lys Cys Thr Ile Lys
Asn Phe Asn Val Ser Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val
Ser Pro Thr Gln Glu Val Ile Arg Phe Pro Asn Ile Thr Asn Arg Cys
Pro Phe Asp Lys Val Phe Asn Ala Thr Arg Phe Pro Asn Val Tyr Ala
Trp Glu Arg Thr Lys Ile Ser Asp Cys Val Ala Asp Tyr Thr Val Leu
Tyr Asn Ser Thr Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro
Ser Lys Leu Ile Asp Leu Cys Phe Thr Ser Val Tyr Ala Asp Thr Phe
Leu Ile Arg Ser Ser Glu Val Arg Gln Val Ala Pro Gly Glu Thr Gly
Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys
Val Ile Ala Trp Asn Thr Ala Lys Gln Asp Gln Gly Gln Tyr Tyr Tyr
Arg Ser His Arg Lys Thr Lys Leu Lys Pro Phe Glu Arg Asp Leu Ser

<400> 2
atgaaaattt taattcttgc tttcctagct agtctagcta aagcacaaga aggatgtggc 60
attatcagtc gaaaacctca gccaaaaatg gcacaagtct cttcttctcg tagaggtgtg 120
tactataatg atgacatttt tcgttctaat gtactacacc tgacgcagga ttatttcctg 180
ccatttgatt caaatttaac acagtacttt tctcttaatg ttgattcaga taggtttacc 240
tactttgaca atcctatttt agactttggt gacggcgtct acttcgctgc tactgaaaag 300
tctaatgtaa ttaggggctg gatttttggt tccactttcg ataacacaac ccagtcagct 360
gttatagtta ataattccac acacattatt atacgtgtgt gcaacttcaa cttatgtaaa 420
gaacctatgt atacagtgtc tcgtggtgca caacaatcat cttgggttta tcagagtgca 480
ttcaattgca catatgatag agtggaaaaa agctttcagc tcgacactgc tcctaaaact 540
ggaaatttta aagacctacg tgagtatgtc tttaagaatc gggatggctt tctcagtgtt 600
taccaaactt atacagctgt taatttacct agaggattac ctattggctt ttcagttttg 660
aggccaattc tcaaactgcc ctttggaatt aacattacat cttatagagt tgttatggct 720
atgtttagcc aaactacttc taatttccta ccagaaagtg ctgcttatta tgttggtaat 780
ttaaaataca ccactttcat gcttagtttt aatgaaaatg ggactattac caatgctatt 840
gattgtgctc aaaacccact tgctgaacta aaatgcacca ttaaaaattt caatgtcagc 900
aagggaatct accaaacatc taacttcaga gtttcgccaa ctcaggaagt tattagattc 960
ccaaacatta caaatcgttg tccttttgac aaagttttta atgctacacg ctttcctaat 1020
gtgtatgcgt gggagagaac taaaatttct gattgtgttg ctgactacac tgttctctac 1080
aactcaactt ctttctcaac ttttaagtgc tatggagttt ctccttctaa gttgattgat 1140
ttatgcttta caagtgtgta tgctgacaca ttcttgataa gatcttcaga agtaagacaa 1200
gttgcaccgg gtgaaactgg tgtcattgct gactataatt acaagctgcc tgatgatttt 1260
actggttgcg taatagcctg gaatactgca aagcaggatc aaggtcagta ttactacagg 1320
tctcaccgga agactaaact taaacctttt gagagagacc tttcttctga tgaaaatggt 1380
gtacgtactc ttagtactta cgacttctac cctagtgtgc cggttgctta tcaggctact 1440
agggtggttg tactgtcatt tgaactacta aacgcacctg caacagtttg tggacctaaa 1500
ttatccacac aacttgttaa gaaccagtgt gtcaatttta attttaatgg actcaaaggt 1560
actggtgttt tgactgaatc atcaaagaga tttcagtcat ttcaacaatt tggtcgtgac 1620
acgtctgatt ttactgactc cgtgcgtgac ccacaaacat tagaaatact tgacatttca 1680
ccatgctctt ttggtggtgt tagtgtaatt acaccaggaa caaatgcttc ttctgaagtg 1740
gctgttcttt atcaagatgt taactgtact gacgtgccag cagcaattca tgcagatcaa 1800
ctaacaccag cttggcgtgt ttattcaact ggaacaaatg ttttccaaac acaggctggc 1860
tgtcttatag gagctgaaca tgttaatgct tcgtatgagt gtgacatccc tattggtgct 1920
ggcatttgtg ctagctacca tacagcttct actttacgta gtgtaggtca gaaatccatt 1980
gtggcttaca ctatgtct 1998

<400> 3
atgaaaattt taattcttgc tttcctagct agtctagcta aagcacaaga aggatgtggc 60
attatcagtc gaaaacctca gccaaaaatg gcacaagtct cttcttctcg tagaggtgtg 120
tactataatg atgacatttt tcgttctaat gtactacacc tgacgcagga ttatttcctg 180
ccatttgatt caaatttaac acagtacttt tctcttaatg ttgattcaga taggtttacc 240
tac 243

<400> 4
tttgacaatc ctattttaga ctttggtgac ggcgtctact tcgctgctac tgaaaagtct 60
aatgtaatta ggggctggat ttttggttcc actttcgata acacaaccca gtcagctgtt 120
atagttaata attccacaca cattattata cgtgtgtgca acttcaactt atgtaaagaa 180
cctatgtata cagtgtctcg tggtgcacaa caatcatctt gggtttatca gagtgcattc 240
aattgcacat atgatagagt ggaaaaaagc tttcagctcg acactgctcc taaaactgga 300
aattttaaag acctacgtga gtatgtcttt aagaatcggg atggctttct cagtgtttac 360
caaacttata cagctgttaa tttacctaga ggattaccta ttggcttttc agttttgagg 420
ccaattctca aactgccctt tggaattaac attacatctt atagagttgt tatggctatg 480
tttagccaaa ctacttctaa tttcctacca gaaagtgctg cttattatgt tggtaattta 540
aaatacacca ctttcatgct tagttttaat gaaaatggga ctattaccaa tgctatt 597

<400> 5
gattgtgctc aaaacccact tgctgaacta aaatgcacca ttaaaaattt caatgtcagc 60
aagggaatct accaaacatc taacttcaga gtttcgccaa ctcaggaagt tattagattc 120
ccaaacatta caaatcgttg tccttttgac aaagttttta atgctacacg ctttcctaat 180
gtgtatgcgt gggagagaac taaaatttct gattgtgttg ctgactacac tgttctctac 240
aactcaactt ctttctcaac ttttaagtgc tatggagttt ctccttctaa gttgattgat 300
ttatgcttta caagtgtgta tgctgacaca ttcttgataa gatcttcaga agtaagacaa 360
gttgcaccgg gtgaaactgg tgtcattgct gactataatt acaagctgcc tgatgatttt 420
actggttgcg taatagcctg gaatactgca aagcaggatc aaggtcagta ttactacagg 480
tctcaccgga agactaaact taaacctttt gagagagacc tttct 525

<400> 6
actgcaaagc aggatcaagg tcagtattac tacaggtctc accggaagac taaacttaaa 60
ccttttgaga gagacctttc ttctgatgaa aatggtgtac gtactcttag tacttacgac 120
ttctacccta gtgtgccggt tgcttatcag gctactaggg tggttgtact gtcatttgaa 180
ctactaaacg cacctgcaac agtttgtgga cctaaattat ccacacaact tgttaagaac 240
cagtgtgtca attttaattt taatggactc aaaggtactg gtgttttgac tgaatcatca 300
aagagatttc agtcatttca acaatttggt cgtgacacgt ctgattttac tgactccgtg 360
cgtgacccac aaacattaga aatacttgac atttcaccat gctcttttgg tggtgttagt 420
gtaattacac caggaaca 438

<400> 7
ccatgctctt ttggtggtgt tagtgtaatt acaccaggaa caaatgcttc ttctgaagtg 60
gctgttcttt atcaagatgt taactgtact gacgtgccag cagcaattca tgcagatcaa 120
ctaacaccag cttggcgtgt ttattcaact ggaacaaatg ttttccaaac acaggctggc 180
tgtcttatag gagctgaaca tgttaatgct tcgtatgagt gtgacatccc tattggtgct 240
ggcatttgtg ctagctacca tacagcttct actttacgta gtgtaggtca gaaatccatt 300
gtggct 306

<400> 8
tgccggatcc attgattgtg ctca 24

<400> 9
cagtgtcgac ttaagaaagg tctctctcaa a 31