Bacillus anthracis
Posted: Wed Oct 10, 2001 7:56 pm
Bacillus anthracis, the organism responsible for the anthrax disease,
carries a plasmid (a free piece of DNA) that codes for three
genes that carry the ultimate toxic disease factors of this organism:
lethal factor (LF), edema factor (EF), and anthrax protective
antigen. In the abstracts obtained from PubMed below, we see research
being carried out creating clones of two of these toxic
factors, which are then introduced into E. coli bacteria. The bacteria
is recultured, and the toxic factors removed, purified, and
tested.
Without pretending to guess from a computer screen whether the intention
of this this type of research is for eventual therapeutic
purposes, one cannot deny the researchers are playing with a very hot
fire. The materials are ultimately no less dangerous than
those used for atomic weapons, or other weapons of mass destruction. It
is time for strict outside oversight of all labs capable of
this type of research, as well as monitoring the movement of key
necessary materials, especially certain organisms supplied by
various companies or research agencies.
At this time when we are again faced with the potential for great
destruction and loss of life, the roots of which lie sometimes
invisible in our midst, please consider contacting your various
officials world-wide to address this problem that may already be out
of control. Even if the possibility of negative intentions are
eliminated, the potential for disastrous accidents of major consequence
due to a natural or man-made disaster, remain an omnipresent concern.
To allow these activities involving genetic manipulation
of micro-organisms to continue without immediate acknowledgement of the
serious dangers involved, is the height of intellectual
arrogance. Please consider what you might do to protect our future, and
that of our children and grandchildren.
----------------------------------------------------------------------------------------------
Enhanced expression of the recombinant lethal factor of Bacillus
anthracis by Fed-Batch culture.
Gupta P, Sahai V, Bhatnagar R.
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi,
110067, India.
....The structural gene for the 90-kDa lethal factor (LF) isolated from
Bacillus anthracis was expressed....in Escherichia coli.
Various strategies were tried to scale up the expression of the
recombinant lethal factor by bioprocess
optimization....The recombinant LF resembled the LF purified from B.
anthracis....It was possible to achieve a yield
of 50 mg of the purified protein from 1 litre culture broth. Copyright
2001 Academic Press.
PMID: 11467855 [PubMed - indexed for MEDLINE]
----------------------------------------------------------------------------
Infect Immun 2001 Oct;69(10):6532-6536
Purification of Anthrax Edema Factor from Escherichia coli and
Identification of Residues Required for Binding to Anthrax Protective
Antigen.
Kumar P, Ahuja N, Bhatnagar R.
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi 110067,
India.
The structural gene for anthrax edema factor (EF) was expressed in
Escherichia coli under the control of a powerful T5 promoter
to yield the 89-kDa recombinant protein that reacted with anti-EF
antibodies. Recombinant EF was purified to homogeneity by a
two-step procedure involving metal chelate affinity chromatography and
cation-exchange chromatography. __ From 1 liter of
culture, 2.5 mg of biologically active EF was easily purified. This is
the first report of purification of anthrax EF from
E. coli. EF purified from E. coli was biologically and functionally as
active as its Bacillus anthracis counterpart.__ The
recombinant protein could compete with lethal factor for binding to
protective antigen. Sequence analysis revealed a stretch of
seven amino acids, Val Tyr Tyr Glu Ile Gly Lys, present both in EF
(residues 136 to 142) and lethal factor (residues 147 to 153). To
investigate the role of these seven residues in binding to protective
antigen, the residues were individually mutated to alanine in
EF. Mutations in residues Tyr137, Tyr138, Ile140, and Lys142 of EF
specifically blocked its interaction with anthrax protective
antigen. The adenylate cyclase activity of the mutants remained
unaffected. The results suggested that residues Tyr137, Tyr138,
Ile140, and Lys142 are required for binding of EF to anthrax protective
antigen, which facilitates its entry into susceptible cells.
PMID: 11553601 [PubMed - in process]
Health, Hope, Joy & Healing :
May you Prosper, even as your Soul Prospers 3John 2
Jennifer Ruby
Email advice is not a substitute for medical treatment.
http://groups.yahoo.com/group/SymphonicHealth
______________________________________________
«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤
¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯
carries a plasmid (a free piece of DNA) that codes for three
genes that carry the ultimate toxic disease factors of this organism:
lethal factor (LF), edema factor (EF), and anthrax protective
antigen. In the abstracts obtained from PubMed below, we see research
being carried out creating clones of two of these toxic
factors, which are then introduced into E. coli bacteria. The bacteria
is recultured, and the toxic factors removed, purified, and
tested.
Without pretending to guess from a computer screen whether the intention
of this this type of research is for eventual therapeutic
purposes, one cannot deny the researchers are playing with a very hot
fire. The materials are ultimately no less dangerous than
those used for atomic weapons, or other weapons of mass destruction. It
is time for strict outside oversight of all labs capable of
this type of research, as well as monitoring the movement of key
necessary materials, especially certain organisms supplied by
various companies or research agencies.
At this time when we are again faced with the potential for great
destruction and loss of life, the roots of which lie sometimes
invisible in our midst, please consider contacting your various
officials world-wide to address this problem that may already be out
of control. Even if the possibility of negative intentions are
eliminated, the potential for disastrous accidents of major consequence
due to a natural or man-made disaster, remain an omnipresent concern.
To allow these activities involving genetic manipulation
of micro-organisms to continue without immediate acknowledgement of the
serious dangers involved, is the height of intellectual
arrogance. Please consider what you might do to protect our future, and
that of our children and grandchildren.
----------------------------------------------------------------------------------------------
Enhanced expression of the recombinant lethal factor of Bacillus
anthracis by Fed-Batch culture.
Gupta P, Sahai V, Bhatnagar R.
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi,
110067, India.
....The structural gene for the 90-kDa lethal factor (LF) isolated from
Bacillus anthracis was expressed....in Escherichia coli.
Various strategies were tried to scale up the expression of the
recombinant lethal factor by bioprocess
optimization....The recombinant LF resembled the LF purified from B.
anthracis....It was possible to achieve a yield
of 50 mg of the purified protein from 1 litre culture broth. Copyright
2001 Academic Press.
PMID: 11467855 [PubMed - indexed for MEDLINE]
----------------------------------------------------------------------------
Infect Immun 2001 Oct;69(10):6532-6536
Purification of Anthrax Edema Factor from Escherichia coli and
Identification of Residues Required for Binding to Anthrax Protective
Antigen.
Kumar P, Ahuja N, Bhatnagar R.
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi 110067,
India.
The structural gene for anthrax edema factor (EF) was expressed in
Escherichia coli under the control of a powerful T5 promoter
to yield the 89-kDa recombinant protein that reacted with anti-EF
antibodies. Recombinant EF was purified to homogeneity by a
two-step procedure involving metal chelate affinity chromatography and
cation-exchange chromatography. __ From 1 liter of
culture, 2.5 mg of biologically active EF was easily purified. This is
the first report of purification of anthrax EF from
E. coli. EF purified from E. coli was biologically and functionally as
active as its Bacillus anthracis counterpart.__ The
recombinant protein could compete with lethal factor for binding to
protective antigen. Sequence analysis revealed a stretch of
seven amino acids, Val Tyr Tyr Glu Ile Gly Lys, present both in EF
(residues 136 to 142) and lethal factor (residues 147 to 153). To
investigate the role of these seven residues in binding to protective
antigen, the residues were individually mutated to alanine in
EF. Mutations in residues Tyr137, Tyr138, Ile140, and Lys142 of EF
specifically blocked its interaction with anthrax protective
antigen. The adenylate cyclase activity of the mutants remained
unaffected. The results suggested that residues Tyr137, Tyr138,
Ile140, and Lys142 are required for binding of EF to anthrax protective
antigen, which facilitates its entry into susceptible cells.
PMID: 11553601 [PubMed - in process]
Health, Hope, Joy & Healing :
May you Prosper, even as your Soul Prospers 3John 2
Jennifer Ruby
Email advice is not a substitute for medical treatment.
http://groups.yahoo.com/group/SymphonicHealth
______________________________________________
«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤»§«¤»¥«¤
¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯