Enzymatic synthesis of uniformly /sup 32/P-labeled polyribonucleotides and high-specific-activity ribonucleoside 5'-alpha-/sup 32/Pdiphosphates

Journal Article · Tue Jan 01 04:00:00 EST 1985 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(85)90119-8· OSTI ID:5613060

Kang, C; Cantor, C R

Uniformly /sup 32/P-labeled polyribonucleotides of high specific activity can be rapidly and easily synthesized from commercially available ribonucleoside 5'-(alpha-/sup 32/P)triphosphates by using two enzymes in sequence. Myosin ATPase completely and irreversibly converted any triphosphates to diphosphates in 10 min. The product diphosphates, without purification, can be polymerized by polynucleotide phosphorylase (PNPase) in 1 h with an average yield of 60%. By choosing the desired molar ratio of radioactive and nonradioactive tri- or diphosphates, polymers of a wide range of specific activity can be obtained. Since myosin ATPase and PNPase both have little base specificity, the method can be used to synthesize a radiolabeled polymer of any desired base composition.

• Research Organization: Columbia Univ., New York, NY
• OSTI ID: 5613060
• Journal Information: Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 144:1; ISSN ANBCA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
ATP-ASE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL PREPARATION
DAYS LIVING RADIOISOTOPES
ENZYME ACTIVITY
ENZYMES
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHOHYDROLASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
RADIOISOTOPES
SYNTHESIS
TRACER TECHNIQUES