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Subject: Global Ti Survey by J. Kirk Wiebe & William (Bill) Binney If you wish to participate in the globaltisurvey, please send your name, address (including postal code & country) & telephone number, email to: globaltisurvey@gmail.com, attn: Kate Ryan/Karla Smith (request a link to the online survey & a code). They are the new scientific group attempt to understand victims claims of electromagnetic assaults and related covert harassment. Must submit by (6/19/2017 – 10/18/2017)
The next two articles are included in their entirety so this information is altogether here to read, excluding the References. Insects have been used to spread Lyme and Morgellons for a reason. Genetic therapy is accomplished by injecting vectors into a person using a needle or perhaps other methods. Genetically altered insects are the needles and can be used to spread genetic changes throughout a population. Hopefully these will help you understand that a bioweapon is being used to make people sick. Its not necessary to murder your enemy when you can disable them and put them out of commission and take their power away without anyone knowing. That is why Morgellons is being denied and called "Delusional Parasitosis". It seems that some people think there are too many of us on the planet and are more than happy to eliminate us with this type of weapon. This was not possible until the invention of a Complete Controllable Inducible Expression System was made available in the mid-90's.
Complete Control Inducible Mammalian Expression System Manual
"The Complete Control inducible mammalian expression system is a gene transfer system that allows precise control of gene expression in a wide variety of mammalian cell types. Development of the Complete Control system is based upon the finding that the insect hormone ecdysone or its analog ponasterone A (ponA) can activate transcription in mammalian cells harboring both the gene for the Drosophila melanogaster ecdysone receptor and a promoter containing a binding site for the ecdysone receptor. The Complete Control system has several advantages over other inducible systems. PonA has no known measurable effect on mammalian physiology. PonA has a short in vivo half-life, and its lipophilic nature allows it to efficiently penetrate all tissues, including the brain. The result is rapid and potent induction of gene expression and rapid clearance. A 1000-fold induction of a reporter gene, with negligible basal expression, has been obtained with the Complete Control system." http://www.genomics.agilent.com/files/Manual/217468.pdf
BIOWEAPONS: Using Expression Vectors to change humans
Expression vector
From Wikipedia, the free encyclopedia
An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the proteinencoded by the gene. Expression vectors are the basic tools in biotechnology for the production of proteins. The vector is engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector. The goal of a well-designed expression vector is the efficient production of protein, and this may be achieved by the production of significant amount of stable messenger RNA, which can then be translated into protein. The expression of a protein may be tightly controlled, and the protein is only produced in significant quantity when necessary through the use of an inducer, in some systems however the protein may be expressed constitutively. Escherichia coli is commonly used as the host for protein production, but other cell types may also be used. An example of the use of expression vector is the production of insulin, which is used for medical treatments of diabetes.
An expression vector has features that any vector may have, such as an origin of replication, a selectable marker, and a suitable site for the insertion of a gene such as the multiple cloning site. The cloned gene may be transferred from a specialized cloning vector to an expression vector, although it is possible to clone directly into an expression vector. The cloning process is normally performed in Escherichia coli, and vectors used for protein production in organisms other than E.coli may have, in addition to a suitable origin of replication for its propagation in E. coli, elements that allow them to be maintained in another organism, and these vectors are called shuttle vectors.
An expression vector must have elements necessary for gene expression. These may include a promoter, the correct translation initiation sequence such as a ribosomal binding site and start codon, a termination codon, and a transcription termination sequence.[2] There are differences in the machinery for protein synthesis between prokaryotes and eukaryotes, therefore the expression vectors must have the elements for expression that is appropriate for the chosen host. For example, prokaryotes expression vectors would have a Shine-Dalgarno sequence at its translation initiation site for the binding of ribosomes, while eukaryotes expression vectors would contain the Kozak consensus sequence.
The promoter initiates the transcription and is therefore the point of control for the expression of the cloned gene. The promoters used in expression vector are normally inducible, meaning that protein synthesis is only initiated when required by the introduction of an inducer such as IPTG. Gene expression however may also be constitutive (i.e. protein is constantly expressed) in some expression vectors. Low level of constitutive protein synthesis may occur even in expression vectors with tightly controlled promoters.
After the expression of the gene product, it is usually necessary to purify the expressed protein; however, separating the protein of interest from the great majority of proteins of the host cell can be a protracted process. To make this purification process easier, a purification tag may be added to the cloned gene. This tag could be histidine (His) tag, other marker peptides, or a fusion partners such as glutathione S-transferase or maltose-binding protein. Some of these fusion partners may also help to increase the solubility of some expressed proteins. Other fusion proteins such as green fluorescent protein may act as a reporter gene for the identification of successful cloned genes.
Others
The expression vector is transformed or transfected into the host cell for protein synthesis. Some expression vectors may have elements for transformation or the insertion of DNA into the host chromosome, for example the vir genes for plant transformation, and integrase sites for chromosomal insertion.
Some vectors may include targeting sequence that may target the expressed protein to a specific location such as the periplasmic space of bacteria.
Expression/Production systems
Different organisms may be used to express a gene's target protein, and the expression vector used will therefore have elements specific for use in the particular organism. The most commonly used organism for protein production is the bacterium Escherichia coli. However, not all proteins can be successfully expressed in E. coli, or be expressed with the correct form of post-translational modifications such as glycosylations, and other systems may therefore be used.
Bacterial
An example of a bacterial expression vector is the pGEX-3x plasmid
The expression host of choice for the expression of many proteins is Escherichia coli as the production of heterologous protein in E. coli is relatively simple and convenient, as well as being rapid and cheap. A large number of E. coli expression plasmids are also available suitable for a wide variety of needs. Other bacteria used for protein production include Bacillus subtilis.
Most heterologous proteins are expressed in the cytoplasm of E. coli. However, not all proteins formed may be soluble in the cytoplasm, and incorrectly folded proteins formed in cytoplasm can form insoluble aggregates called inclusion bodies. Such insoluble proteins will require refolding, which can be an involved process and may not necessarily produce high yield.[3] Proteins which have disulphide bonds are often not able to fold correctly due to the oxidizing environment in the cytoplasm which prevents such bond formation, and a possible solution is to target the protein to the periplasmic space by the use of an N-terminal signal sequence. Other more sophisticated systems are also being developed; such systems may allow for the expression of proteins previously thought impossible in E. coli, such as glycosylated proteins.[4][5]
The promoters used for these vector are usually based on the promoter of the lac operon or the T7 promoter,[6] and they are normally regulated by the lacoperator. These promoters may also be hybrids of different promoters, for example, the Tac-Promoter is a hybrid of trp and lac promoters.[7] Note that most commonly used lac or lac-derived promoters are based on the lacUV5 mutant which is insensitive to catabolite repression. This mutant allows for expression of protein under the control of the lac promoter when the growth medium contains glucose since glucose would inhibit gene expression if wild-type lac promoter is used.[8] Presence of glucose nevertheless may still be used to reduce background expression through residual inhibition in some systems.[9]
Examples of E. coli expression vectors are the pGEX series of vectors where glutathione S-transferase is used as a fusion partner and gene expression is under the control of the tac promoter,[10][11][12] and the pET series of vectors which uses a T7 promoter.[13]
It is possible to simultaneously express two or more different proteins in E. coli using different plasmids. However, when 2 or more plasmids are used, each plasmid needs to use a different antibiotic selection as well as a different origin of replication, otherwise the plasmids may not be stably maintained. Many commonly used plasmids are based on the ColE1 replicon and are therefore incompatible with each other; in order for a ColE1-based plasmid to coexist with another in the same cell, the other would need to be of a different replicon, e.g. a p15A replicon-based plasmid such as the pACYC series of plasmids.[14] Another approach would be to use a single two-cistron vector or design the coding sequences in tandem as a bi- or poly-cistronic construct.[15][16]
Yeast
A yeast commonly used for protein production is Pichia pastoris.[17] Examples of yeast expression vector in Pichia are the pPIC series of vectors, and these vectors use the AOX1 promoter which is inducible with methanol.[18] The plasmids may contain elements for insertion of foreign DNA into the yeast genome and signal sequence for the secretion of expressed protein. Proteins with disulphide bonds and glycosylation can be efficiently produced in yeast. Another yeast used for protein production is Kluyveromyces lactis and the gene is expressed, driven by a variant of the strong lactase LAC4 promoter.[19]
Saccharomyces cerevisiae is particularly widely used for gene expression studies in yeast, for example in yeast two-hybrid system for the study of protein-protein interaction.[20] The vectors used in yeast two-hybrid system contain fusion partners for two cloned genes that allow the transcription of a reporter gene when there is interaction between the two proteins expressed from the cloned genes.
Baculovirus
Baculovirus, a rod-shaped virus which infect insect cells, is used as the expression vector in this system. Insect cell lines derived from Lepidopterans (moths and butterflies), such as Spodoptera frugiperda, are used as host. The shuttle vector is called bacmid, and gene expression is under the control of a strong promoter pPolh.[21] Baculovirus has also been used with mammalian cell lines in the BacMam system.[22]
Baculovirus is normally used for production of glycoproteins, although the glycosylations may be different from those found in vertebrates. In general, it is safer to use than mammalian virus as it has a limited host range and does not infect vertebrates without modifications.
Plant
Many plant expression vectors are based on the Ti plasmid of Agrobacterium tumefaciens.[23] In these expression vectors, DNA to be inserted into plant is cloned into the T-DNA, a stretch of DNA flanked by a 25-bp direct repeat sequence at either end, and which can integrate into the plant genome. The T-DNA also contains the selectable marker. The Agrobacterium provides a mechanism for transformation, integration of into the plant genome, and the promoters for its vir genes may also be used for the cloned genes. Concerns over the transfer of bacterial or viral genetic material into the plant however have led to the development of vectors called intragenic vectors whereby functional equivalents of plant genome are used so that there is no transfer of genetic material from an alien species into the plant.[24]
Plant viruses may be used as vectors since the Agrobacterium method does not work for all plants. Examples of plant virus used are the tobacco mosaic virus (TMV), potato virus X, and cowpea mosaic virus.[25] The protein may be expressed as a fusion to the coat protein of the virus and is displayed on the surface of assembled viral particles, or as an unfused protein that accumulates within the plant. Expression in plant using plant vectors is often constitutive,[26] and a commonly used constitutive promoter in plant expression vectors is the cauliflower mosaic virus (CaMV) 35S promoter.[27][28]
Mammalian
Mammalian expression vectors offer considerable advantages for the expression of mammalian proteins over bacterial expression systems - proper folding, post-translational modifications, and relevant enzymatic activity. It may also be more desirable than other eukaryotic non-mammalian systems whereby the proteins expressed may not contain the correct glycosylations. It is of particular use in producing membrane-associating proteins that require chaperones for proper folding and stability as well as containing numerous post-translational modifications. The downside, however, is the low yield of product in comparison to prokaryotic vectors as well as the costly nature of the techniques involved. Its complicated technology, and potential contamination with animal viruses of mammalian cell expression have also placed a constraint on its use in large-scale industrial production.[29]
Cultured mammalian cell lines such as the Chinese hamster ovary (CHO), COS, including human cell lines such as HEK and HeLa may be used to produce protein. Vectors are transfected into the cells and the DNA may be integrated into the genome by homologous recombination in the case of stable transfection, or the cells may be transiently transfected. Examples of mammalian expression vectors include the adenoviral vectors,[30] the pSV and the pCMV series of plasmid vectors, vaccinia and retroviral vectors,[31] as well as baculovirus.[22] The promoters for cytomegalovirus (CMV) and SV40 are commonly used in mammalian expression vectors to drive gene expression. Non-viral promoter, such as the elongation factor (EF)-1 promoter, is also known.[32]
Cell-free systems
E. colicell lysate containing the cellular components required for transcription and translation are used in this in vitro method of protein production. The advantage of such system is that protein may be produced much faster than those produced in vivo since it does not require time to culture the cells, but it is also more expensive. Vectors used for E. coli expression can be used in this system although specifically designed vectors for this system are also available. Eukaryotic cell extracts may also be used in other cell-free systems, for example, the wheat germ cell-free expression systems.[33] Mammalian cell-free systems have also been produced.[34]
Applications
Laboratory use
Expression vector in an expression host is now the usual method used in laboratories to produce proteins for research. Most proteins are produced in E. coli, but for glycosylated proteins and those with disulphide bonds, yeast, baculovirus and mammalian systems may be used.
Production of peptide and protein pharmaceuticals
Most protein pharmaceuticals are now produced through recombinant DNA technology using expression vectors. These peptide and protein pharmaceuticals may be hormones, vaccines, antibiotics, antibodies, and enzymes.[35] The first human recombinant protein used for disease management, insulin, was introduced in 1982.[35] Biotechnology allows these peptide and protein pharmaceuticals, some of which were previously rare or difficult to obtain, to be produced in large quantity. It also reduces the risks of contaminants such as host viruses, toxins and prions. Examples from the past include prion contamination in growth hormone extracted from pituitary glands harvested from human cadavers, which caused Creutzfeldt–Jakob disease in patients receiving treatment for dwarfism,[36] and viral contaminants in clotting factor VIII isolated from human blood that resulted in the transmission of viral diseases such as hepatitis and AIDS.[37][38] Such risk is reduced or removed completely when the proteins are produced in non-human host cells.
Transgenic plant and animals
Genetically engineered fluorescent GloFish
In recent years, expression vectors have been used to introduce specific genes into plants and animals to produce transgenic organisms, for example in agriculture it is used to produce transgenic plants. Expression vectors have been used to introduce a vitamin A precursor, beta-carotene, into rice plants. This product is called golden rice. This process has also been used to introduce a gene into plants that produces an insecticide, called Bacillus thuringiensis toxin or Bt toxin which reduces the need for farmers to apply insecticides since it is produced by the modified organism. In addition expression vectors are used to extend the ripeness of tomatoes by altering the plant so that it produces less of the chemical that causes the tomatoes to rot.[39] There have been controversies over using expression vectors to modify crops due to the fact that there might be unknown health risks, possibilities of companies patenting certain genetically modified food crops, and ethical concerns. Nevertheless, this technique is still being used and heavily researched. Transgenic animals have also been produced to study animal biochemical processes and human diseases, or used to produce pharmaceuticals and other proteins. They may also be engineered to have advantageous or useful traits. Green fluorescent protein is sometimes used as tags which results in animal that can fluoresce, and this have been exploited commercially to produce the fluorescent GloFish.
Gene therapy is a promising treatment for a number of diseases where a "normal" gene carried by the vector is inserted into the genome, to replace an "abnormal" gene or supplement the expression of particular gene. Viral vectors are generally used but other nonviral methods of delivery are being developed. The treatment is still a risky option due to the viral vector used which can cause ill-effects, for example giving rise to insertional mutation that can result in cancer.[40][41] However, there have been promising results.[42]
Genome sequencing has given rise to a new generation of genetically engineered bioweapons carrying the potential to change the nature of modern warfare and defense.
Introduction Biological weapons are designed to spread disease among people, plants, and animals through the introduction of toxins and microorganisms such as viruses and bacteria. The method through which a biological weapon is deployed depends on the agent itself, its preparation, its durability, and the route of infection. Attackers may disperse these agents through aerosols or food and water supplies (1).
Although bioweapons have been used in war for many centuries, a recent surge in genetic understanding, as well as a rapid growth in computational power, has allowed genetic engineering to play a larger role in the development of new bioweapons. In the bioweapon industry, genetic engineering can be used to manipulate genes to create new pathogenic characteristics aimed at enhancing the efficacy of the weapon through increased survivability, infectivity, virulence, and drug resistance (2). While the positive societal implications of improved biotechnology are apparent, the “black biology” of bioweapon development may be “one of the gravest threats we will face” (2).
Limits of Past Bioweapons Prior to recent advances in genetic engineering, bioweapons were exclusively natural pathogens. Agents must fulfill numerous prerequisites to be considered effective military bioweapons, and most naturally occurring pathogens are ill suited for this purpose (3). First, bioweapons must be produced in large quantities. A pathogen can be obtained from the natural environment if enough can be collected to allow purification and testing of its properties. Otherwise, pathogens could be produced in a microbiology laboratory or bank, a process which is limited by pathogen accessibility and the safety with which the pathogens can be handled in facilities. To replicate viruses and some bacteria, living cells are required. The growth of large quantities of an agent can be limited by equipment, space, and the health risks associated with the handling of hazardous germs (1). In addition to large-scale production, effective bioweapons must act quickly, be environmentally robust, and their effects must be treatable for those who are implementing the bioweapon (3).
Recent Advances As researchers continue to transition from the era of DNA sequencing into the era of DNA synthesis, it may soon become feasible to synthesize any virus whose DNA sequence is known (4). This was first demonstrated in 2001 when Dr. Eckard Wimmer re-created the poliovirus and again in 2005 when Dr. Jeffrey Taubenberger and Terrence Tumpey re-created the 1918 influenza virus (1). The progress of DNA synthesis technology will also allow for the creation of novel pathogens. According to biological warfare expert Dr. Steven Block, genetically engineered pathogens “could be made safer to handle, easier to distribute, capable of ethnic specificity, or be made to cause higher mortality rates” (2).
The growing accessibility of DNA synthesis capabilities, computational power, and information means that a growing number of people will have the capacity to produce bioweapons. Scientists have been able to transform the four letters of DNA—A (adenine), C (cytosine), G (guanine), and T (thymine)—into the ones and zeroes of binary code. This transformation makes genetic engineering a matter of electronic manipulation, which decreases the cost of the technique (4). According to former Secretary of State Hillary Clinton, “the emerging gene synthesis industry is making genetic material more widely available […] A crude but effective terrorist weapon can be made using a small sample of any number of widely available pathogens, inexpensive equipment, and college-level chemistry and biology.” (5)
Techniques to Enhance Efficacy of Bioweapons Scientists and genetic engineers are considering several techniques to increase the efficacy of pathogens in warfare.
1. Binary Biological Weapons This technique involves inserting plasmids, small bacterial DNA fragments, into the DNA of other bacteria in order to increase virulence or other pathogenic properties within the host bacteria (2).
2. Designer Genes According to the European Bioinformatics Institute, as of December 2012, scientists had sequenced the genomes of 3139 viruses, 1016 plasmids, and 2167 bacteria, some of which are published on the internet and are therefore accessible to the public (6). With complete genomes available and the aforementioned advances in gene synthesis, scientists will soon be able to design pathogens by creating synthetic genes, synthetic viruses, and possibly entirely new organisms (2).
3. Gene Therapy Gene therapy involves repairing or replacing a gene of an organism, permanently changing its genetic composition. By replacing existing genes with harmful genes, this technique can be used to manufacture bioweapons (2).
4. Stealth Viruses Stealth viruses are viral infections that enter cells and remain dormant for an extended amount of time until triggered externally to cause disease. In the context of warfare, these viruses could be spread to a large population, and activation could either be delayed or used as a threat for blackmail (2).
5. Host-Swapping Diseases Much like the naturally occurring West Nile and Ebola viruses, animal viruses could potentially be genetically modified and developed to infect humans as a potent biowarfare tactic (2).
6. Designer Diseases Biotechnology may be used to manipulate cellular mechanisms to cause disease. For example, an agent could be designed to induce cells to multiply uncontrollably, as in cancer, or to initiate apoptosis, programmed cell death (2).
7. Personalized Bioweapons In coming years it may be conceivable to design a pathogen that targets a specific person’s genome. This agent may spread through populations showing minimal or no symptoms, yet it would be fatal to the intended target (4).
Biodefense In addition to creating bioweapons, the emerging tools of genetic knowledge and biological technology may be used as a means of defense against these weapons.
1. Human Genome Literacy As scientific research continues to reveal the functions of specific genes and how genetic components affect disease in humans, vaccines and drugs can be designed to combat particular pathogens based on analysis of their particular molecular effect on the human cell (2).
2. Immune System Enhancement In addition to enabling more effective drug development, human genome literacy allows for a better understanding of the immune system. Thus, genetic engineering can be used to enhance human immune response to pathogens. As an example, Dr. Ken Alibek is conducting cellular research in pursuit of protection against the bioweapon anthrax (2).
3. Viral and Bacterial Genome Literacy Decoding the genomes of viruses and bacteria will lead to molecular explanations behind virulence and drug resistance. With this information, bacteria can be engineered to produce bioregulators against pathogens. For example, Xoma Corporation has patented a bactericidal/permeability-increasing (BPI) protein, made from genes inserted into bacterial DNA, which reverses the resistance characteristic of particular bacteria against some popular antibiotics (2).
4. Efficient Bio-Agent Detection and Identification Equipment Because the capability of comparing genomes using DNA assays has already been acquired, such technology may be developed to identify pathogens using information from bacterial and viral genomes. Such a detector could be used to identify the composition of bioweapons based on their genomes, reducing present-day delays in resultant treatment and/or preventive measures (2).
5. New Vaccines Current scientific research projects involve genetic manipulation of viruses to create vaccines that provide immunity against multiple diseases with a single treatment (2).
6. New Antibiotics and Antiviral Drugs Currently, antibiotic drugs target DNA synthesis, protein synthesis, and cell-wall synthesis processes in bacterial cells. With an increased understanding of microbial genomes, other proteins essential to bacterial viability can be targeted to create new classes of antibiotics. Eventually, broad-spectrum, rather than protein-specific, anti-microbial drugs may be developed (2).
Future of Warfare “The revolution in molecular biology and biotechnology can be considered as a potential Revolution of Military Affairs (RMA),” states Colonel Michael Ainscough, MD, MPH (2). According to Andrew Krepinevich, who originally coined the term RMA, “technological advancement, incorporation of this new technology into military systems, military operational advancement, and organizational adaptation in a way that fundamentally alters the character and conduct of conflict” are the four components that make up an RMA. For instance, the Gulf War has been classified as the beginning of the space information warfare RMA. “From the technological advances in biotechnology, biowarfare with genetically engineered pathogens may constitute a future such RMA,” says Ainscough (2).
In addition, the exponential increase in computational power combined with the accessibility of genetic information and biological tools to the general public and lack of governmental regulation raise concerns about the threat of biowarfare arising from outside the military (7). The US government has cited the efforts of terrorist networks, such as al Qaida, to recruit scientists capable of creating bioweapons as a national security concern and “has urged countries to be more open about their efforts to clamp down on the threat of bioweapons” (5).
Despite these efforts, biological research that can potentially lead to bioweapon development is “far more international, far more spread out, and far more diverse than nuclear science […] researchers communicate much more rapidly with one another by means that no government can control […] this was not true in the nuclear era,” according to David Kay, former chief U.S. weapons inspector in Iraq (7). Kay is “extraordinarily pessimistic that we [the United States] will take any of the necessary steps to avoid the threat of bioweapons absent their first actual use” (7).
“There are those who say: ‘the First World War was chemical; the Second World War was nuclear; and that the Third World War – God forbid – will be biological’” (2).
ACTIVIST: Mike Mason
ACTIVIST ELLA
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TI'S IN CHINA
An Announcement To The Whole World By Chinese Victims Of Electronic And Psychotronic Weapons
A Linqstat order is here. If you are interested, please email Neal to order. electricrose22@yahoo.com
If you have a tent, you can just tape some pieces of Linqstat together and drape them over the tent. You could make it the exact shape as the rain cover. You can ground it to an outlet, to a ground with a banana clip or attach a TENS electrode to it to create an energy field through the material. Using a tent structure that's already got places to attach the Linqstat make it a lot easier. You can even get a simple one and put it on your bed to drape the Lingstat over. Completely covering an already-made tent like this one with Linqstat is pretty easy and you can just put the TENS pad on the outside of it and "light it up", keeping the frequencies on the outside.
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Genetically Engineered Bioweapons: A New Breed of Weapons for Modern Warfare
You can even get a simple one and put it on your bed to drape the Lingstat over. Completely covering an already-made tent like this one with Linqstat is pretty easy and you can just put the TENS pad on the outside of it and "light it up", keeping the frequencies on the outside.