Efficient template-free enzymatic synthesis of polynucleotides

Efficient template-free enzymatic synthesis of polynucleotides

The present invention relates to compositions and methods for the enzymatic template-free synthesis of polynucleotides wherein different 3 '-O-reversibly blocked deoxynucleoside triphosphates (dNTPs) are incorporated into the growth chain using different terminal deoxynucleotidyl transferase (T...

Ausführliche Beschreibung

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Bibliographische Detailangaben
Hauptverfasser: SOSKIN MIKHAIL, SUNI ELODIE, GODREN XAVIER, CHAMPION ELISE, YBERT THOMAS, HEINISCH TILMAN
Format: Patent
Sprache:chi ; eng
Schlagworte:
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Beschreibung
Zusammenfassung:The present invention relates to compositions and methods for the enzymatic template-free synthesis of polynucleotides wherein different 3 '-O-reversibly blocked deoxynucleoside triphosphates (dNTPs) are incorporated into the growth chain using different terminal deoxynucleotidyl transferase (TdT) variants. In part, the present invention recognizes and understands that different TdT variants can be designed to preferentially incorporate a specific dNTP at a higher efficiency than a single "universal" TdT variant for incorporating all four 3 '-O-reversibly blocked dNTPs. 本发明涉及多核苷酸的酶促无模板合成的组合物和方法，其中使用不同的末端脱氧核苷酸转移酶(TdT)变体将不同的3'-O-可逆封闭的脱氧核苷三磷酸(dNTP)掺入到生长链中。部分地，本发明认识和理解到，可以设计不同的TdT变体，以比单一"通用"TdT变体更高的效率优先掺入特异性dNTP，所述单一"通用"TdT变体用于掺入所有四种3'-O-可逆封闭的dNTP。