The active and inactive structures of the Escherichia coli cAMP receptor protein (CRP), a model bacterial transcr!ption
factor, are compared to generate a paradigm in the cAMP-induced activation of CRP. The resulting paradigm is shown to be
consistent with numerous biochemical studies of CRP and CRP*, a group of CRP mutants displaying cAMP-free activity.
The cAMP affinity of CRP is dictated by two factors: (i) the effectiveness of the cAMP pocket and (ii) the protein equilibrium
of apo-CRP. How these two factors interplay in determining the cAMP affinity and cAMP specificity of CRP and CRP*
mutants are discussed. Both the current understanding and knowledge gaps of CRP-DNA interactions are also described.
This review ends with a list of several important CRP issues that need to be addressed in the future.
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Various treatments and agents had been reported to inactivate
RNA viruses. Of these, thermal inactivation is generally
considered an effective and cheap method of sample
preparation for downstream assays. The purpose of this study
is to establish a safe inactivation method for SARS-CoV-2
without compromising the amount of amplifiable viral genome
necessary for clinical diagnoses. In this study, we demonstrate
the infectivity and genomic stability of SARSCoV-
2 by thermal inactivation at both 56°C and 65°C. The results substantiate that viable SARS-CoV-2 is readily inactivated
when incubated at 56°C for 30 min or at 65°C for
10 min. qRT-PCR of specimens heat-inactivated at 56°C for
30 min or 65°C for 15 min revealed similar genomic RNA
stability compared with non-heat inactivated specimens. Further,
we demonstrate that 30 min of thermal inactivation at
56°C could inactivate viable viruses from clinical COVID-19
specimens without attenuating the qRT-PCR diagnostic sensitivity.
Heat treatment of clinical specimens from COVID-19
patients at 56°C for 30 min or 65°C for 15 min could be a useful method for the inactivation of a highly contagious agent,
SARS-CoV-2. Use of this method would reduce the potential
for secondary infections in BSL2 conditions during diagnostic
procedures. Importantly, infectious virus can be inactivated
in clinical specimens without compromising the
sensitivity of the diagnostic RT-PCR assay.
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