Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1995-05-08
1999-01-19
McElwain, Elizabeth
Chemistry: molecular biology and microbiology
Spore forming or isolating process
435 691, 4351723, 4352523, 4353201, 800205, 536 232, 536 235, A01H 500, C12N 922, C12N 1512, C12N 1582
Patent
active
058613005
ABSTRACT:
Isolated 2-5A-dependent RNases, an interferon-induced enzyme which is activated by 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A) and implicated in both the molecular mechanisms of interferon action and in the fundamental control of RNA stability in mammalian cells, and encoding sequences therefor are disclosed. The expression cloning and analysis of murine and human 2-5A-dependent RNases is also disclosed. Recombinant human 2-5A-dependent RNase produced in vitro bound an activating affinity matrix, 2-5A-cellulose, resulting in ribonuclease activity. The 2-5A binding properties of the recombinant and naturally occurring forms of 2-5A-dependent RNase are basically identical. Interferon induction of 2-5A-dependent RNase expression is demonstrated by measuring the mRNA levels in cells treated with interferon and cycloheximide. Analysis of aligned murine and human 2-5A-dependent RNase sequences revealed several features, including similarity to RNase E which is implicated in the control of mRNA stability in E. coli. A duplicated phosphate-binding loop motif is determined by deletion analysis and site-directed mutagenesis to function in the binding of 2-5A. In addition, recombinant nucleotide sequences, recombinant vectors, recombinant cells and antiviral plants which express, for example, amino acid sequences which have activity that interfere with or inhibit viral replication are disclosed.
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SenGupta Dibyendu N.
Silverman Robert H.
McElwain Elizabeth
The Cleveland Clinic Foundation
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