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Related post: - Moss 15-19
Expression of Genes of Respiratory Viruses by
Vaccinia Virus Recombinants - Moss 15-20
(Incorporated into ZOl AI 00298-07)
Expression of Hepatitis B Virus Genes by Vaccina
Virus Recombinants - Moss 15-21
(Incorporated into ZOl AI 00298-07)
Expression of Malaria Genes by Vaccirua Virus
Recombinants - Moss 15-22
(Incorporated into ZOl AI 00298-07)
Recombinant Vaccines Against Retroviruses Associated
With Leukemia and AIDS - Moss 15-23
Vaccine Virus Growth Factor - Buller 15-24
i
ZOl AI 00445-04 LVD
ZOl AI 00538-01 LVD
ZOl AI 00539-01 LVD
ZOl AI OOStO-01 LVD
ZOl AI 00541-01 LVD
ZOl AI 00542-01 LVD
Mechanism ot Viral DN'A Replication - Challberg . 15-25
Interaction of Human Immunodeficiency Virus with
CD4 Receptor - Moss 15-26
Virus-cell Interactioris, Viral Pathogenesis and
Host Immunity - Moss 15-27
Molecular Biology of Human Herpesviruses
- Frenkel 15-28
Folding Assembly, and Trar\sport of Viral
Glycoproteins - Yewdell 15-29
Processing of Viral Proteins for T Cell Recognition
- Bennink 15-30
Public Health Service-National Institutes of Health
ANNUAL REPORT OF THE LABORATORY OF VIRAL DISEASES, NIAID
October 1, 1987 to September 30, 1988
Dr. Bernard Moss
Chief, Laboratory of Viral Diseases
RESEARCH OBJECTIVES
The Laboratory of Viral Diseases carries out investigations on the molecular
biology of viruses, the interactions of viruses with host cells, the pathogenesis of viral
diseases, and host defense mechanisms that are designed to increase fundamental
knowledge and facilitate the development of new approaches to the prevention and
treatment of virus infections. Current topics of basic research include: regulation of gene
expression, mechanisms of DNA replication, assembly and transport of virus particles,
virus growth factors, determinants of Tadora 20 Mg virus virulence, host resistance genes, and viral
targets of humoral and cell mediated immunity. More applied areas of investigation
include the development of recombinant expression vectors, candidate vaccines, and
antiviral agents. These studies involve a wide range of viruses including human
immunodeficiency virus.
REGULATION OF VIRUS GENE EXPRESSION
DNA viruses provide model systems for studying basic aspects of gene expression
and can be used as expression vectors for a variety of purposes. Several different virus
systems are currently under investigation by members of LVD.
POXVIRUSES
Poxviruses are large DNA viruses that replicate in the cytoplasm of infected cells.
In order to express and replicate their genomes outside of the nucleus, poxviruses must
encode enzymes for these processes. This feature makes poxviruses, among which
vaccinia virus is the prototype, unique for studying the regulation of transcription.
Additional information regarding transcription is also needed to improve vaccinia virus
expression vectors which are currently being tested as candidate live Buy Tadora vaccines against
several diseases including AIDS. EXinng the past year progress has been made m
determining the signals used for irutiation and termination of transcription of early and
late vaccinia virus genes and the enzymes Tadora Online and factors needed to carry out these steps.
Purification and characterization of a factor required for transcription of vaccinia
virus early genes . The inability of highly purified vaccinia virus RNA polymerase to
transcribe vaccinia virus genes led to our discovery of a specific transcription factor. A
complementation assay was developed to quantitate the recovery of the factor during
purification. An observation that the factor binds specfically to early vaccinia virus
promoters was exploited in the final affinity chromatography step. The purified protein,
which we have named VETF, has a molecular weight of approximately 150,000 and
contains two polypeptides of 77,000 and 82,000. The purified factor stimulates
trar\scription from all early promoters tested, binds specifically to early promoters, and
exhibits a DNA-dependent ATPase activity. The latter may explain the previously
described requirement for ATP hydrolysis in transcription. We suspect that VETF is the
functional equivalent of the eukaryotic TATA binding factor which has not yet been
purified. (Broyles, Shuman and Moss)
15-1
Factor-dependent transcription termination bv vnccinia RNA polymernse . The
mechanism used tor termination ot mR\'A in eukaryotic cells is not yet known.
Transcription termination by vaconia RNA polymerase is dependent on a trans-acting
factor that we have named VTF and which is associated witn the vaccinia capping
enzyme. VTF induces termination approximately 50 nucleotides downstream ot a signal
sequence TTTTTNT in the template strand. Since the signal is transcribed, we carried
out experiments to determine tne role of the nascent RNA in termination. We found
that halogenated analogs of LTP specifically abrogated factor-dependent termination
without affecting the synthesis of read-through transcripts. These results indicate that
the cis-acting termination signal is recognized in the nascent RNA rather than the
template DNA.
(Shuman and Moss).
Late vaccinia virus mRNA has capped polv(A) leaders of variable length . Several
research groups have recently obtained evidence for a novel poly(A) structure at the 5'
ends of vaccinia virus late n\RNAs. We have demonstrated capped poly(A) leaders of
variable length located immediately upstream of the translation initiation codon by
direct analysis of a major late RNA species. A "decapping-recapping" protocol was used
to specdficaUv substitute a radioactively labeled phosphate for an unlabeled one within
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