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  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
  • C07C227/30—Preparation of optical isomers
  • C07C227/34—Preparation of optical isomers by separation of optical isomers
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
  • C07C227/38—Separation; Purification; Stabilisation; Use of additives
  • C07C227/40—Separation; Purification
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C273/00—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
  • C07C273/18—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups of substituted ureas
  • C07C273/189—Purification, separation, stabilisation, use of additives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C277/00—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
  • C07C277/08—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups of substituted guanidines
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
  • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
  • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
  • C12Q1/6811—Selection methods for production or design of target specific oligonucleotides or binding molecules

Definitions

• This invention relates to isolated RNA molecules with high or
• the invention are processes lor securing these RNA molecules.
• a third example is the interaction of the H ⁇ V-1 TAT protein with a stem loop structure of TAR RNA, located
• TAR is a single arginine within a basic region of TAT.
• RNA to largely determine specificity, functionality, and
• RNA aptamers which specifically recognize amino acids
• arginine specific aptamers might be especially relevant to protein-RNA
• the HIV-1 Rev protein contains a basic region in which ten of seventeen amino acids between positions
• RRE responsive element
• TAR/ Tat complex an -irginine and an isoleucine residue were found to
• Rex-protein of HTLV-I might interact with its natural
• the present invention is a
• Figure 1 is a representative elution protocol obtained in
• the first pool (“pool 1" hereafter) was
• pool 2 was then subjected to PCR amplification.
• the PCR amplification was carrier out as described by Famulok, supra,
• amplification product was extracted with phenol/ CHCL y precipitated with ethanol, dissolved in 1.3 ml of TE (lOmM Tris-HCl, pH 7.6; ImM
• RNA produced in Example 1, supra was selected for high
• the preselection column consisted of
• RNA containing samples were added to the preselection column, and then washed with 2-3 volumes of buffer. The effect of this
• the column was then eluted with another 5 column volumes of binding buffer containing 20mM citrulline so as to remove any non-
• binding buffer plus 20mM arginine was then heat denatured, in
• RNA typically
• RNA molecules were used, i.e., Ag. 06 (SEQ ID NO: ), as well
• the value for Ag. 06 was about 330nM, i.e., nearly 200 times greater
• aminoglycoside antibiotics tobramycin, kanamycin, lividomycin
• RNA ribonucleic acid
• isolated ribonucleic acid molecules having
• affinities of around 300nM, more preferably greater than about 60 ⁇ M are affinities of around 300nM, more preferably greater than about 60 ⁇ M.
• RNA molecules can have the strong affinity for any L- or D-
• RNA molecules are those set forth in SEQ ID NOS: 3 through 20 , inclusive.
• RNA molecules have various uses, which will be clear to
• RNA molecules may be used to
• affinitv to form complexes of RNA and amino acid which can be
• Enantiomers of amino acids may be identified and/or
• amino acids such as L-arginine and L- ⁇ trulline may be separated, and
• the bound RNA is contacted with a solution of an amino acid which is not the amino acid for which a high affinity binder is
• steps using L-citrulline Preferably, one carries out the cycling for
• affinity binder is sought. This serves to elute the desired RNA from
• RNA be bound to the RNA
• a strong or high affinity binder as was noted, supra, is an RNA molecule which has a low Kd value.
• strong affinitv (or high affinity) binders are those with a Kd
• these molecules have a Kd
• this value is about 500 nM or less, and most

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• General Engineering & Computer Science (AREA)
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• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

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• 1997
  • 1997-03-11 EP EP97915359A patent/EP0888367A1/en not_active Withdrawn
  • 1997-03-11 WO PCT/EP1997/001223 patent/WO1997033895A1/en not_active Application Discontinuation
  • 1997-03-11 JP JP9532275A patent/JP2000507097A/ja active Pending

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