[Microbiology] Request :- Can anyone perform this experiment?

jitesh dundas via microbio%40net.bio.net (by jbdundas from gmail.com)
Fri Dec 11 09:05:38 EST 2009


Dear All,

Can anyone perform this experiment for me. I will add him/her as a co-author
to my paper on this experiment.

Details:-
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Experiment to be carried out
Materials and Methods:
T-Antigen which is an antigen that requires the participation of T
lymphocytes even before an immune response could occur. As most antigens are
of this type, the antigen that requires the interaction between T and B
cells will be used to initiate antibody production. Viral stocks for the
analysis will be generated by transient transfection of 293T cells [Fouchier
RA et al 1997]. The virus will then be quantified by an enzyme-linked
immunosorbent assay (ELISA) for identifying the soluble concentration. In
order to generate high-titre viruses, the  293T cells can first transiently
be cotransfected with other virions [needs to be indentified and reviewed
carefully]. After infection, the input virus will be  removed by extensive
washing with phosphate-buffered saline (PBS), and the T cells would be
incubated in fresh media for 36 hours to allow viral production to reach a
high level. The cells will then washed again with PBS and placed in the
appropriate medium for viral production. Further post-production, the
virus-containing supernatants would be centrifuged for 5 min at 500 x g and
filtered through 0.45-µm-pore-size filters. The infectivity of the viruses
will be determined by infection  using logarithmic time scale. The T-Cells
within multicellular biological systems were infected with viruses in
different conditions and methods. There will also be several cases executed
based on which experiments will be conducted on cells.  When an unplanned
event occurs with a cell such as pathogen infiltration, or high temperature,
etc then a cell is able to respond in a particular behaviour. Here we will
compare the human body (multi-cellular organism) and the cell (single cell
of human body) with their behaviour capabilities & try to answer the
questions in our work favourably. The behavioural aspects of a cell may also
be studied to understand more about them. Their vulnerabilities and coping
mechanisms could give us an idea of how cells resist diseases. In the
experiments mentioned in the methodology, the test cases may be observed or
improved to study the movements (or changes in the cell behaviour) in
different conditions. For example, how does a cell react when it is invaded
by a pathogen? what are the changes that happen to its structure and
functionality? Any such changes or irregularity can be studied here. We
believe comparing the test cases will reveal interesting observation about
the cell.


Reference:Fouchier, R. A., B. E. Meyer, J. H. Simon, U. Fischer, and M. H.
Malim. 1997. HIV-1 infection of non-dividing cells: evidence that the
amino-terminal basic region of the viral matrix protein is important for Gag
processing but not for post-entry nuclear import. EMBO J. 16:4531-4539
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Regards,
Jitesh Dundas
+91-9561527190 / +91-9860925706


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