Recombinant protein structural domain enhanced TET enzyme and whole genome DNA methylation detection method

The invention provides a recombinant protein structural domain for enhancing TET enzyme activity, the recombinant protein structural domain is a DNA methylation binding structural domain (MBD), and the amino acid sequence of MBD1-4 is as shown in SEQ ID 1-4. The recombinant protein MBD-TET formed by...

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Hauptverfasser:	CAO ZHEN, SONG DONGLIANG, HOU CE, WEI LEI, TENG YIGANG, CHEN JINGJING, HUANG KAIYU, JIANG AO
Format:	Patent
Sprache:	chi ; eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMBINATORIAL CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING ORGANIC CHEMISTRY PEPTIDES PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Beschreibung
Zusammenfassung:	The invention provides a recombinant protein structural domain for enhancing TET enzyme activity, the recombinant protein structural domain is a DNA methylation binding structural domain (MBD), and the amino acid sequence of MBD1-4 is as shown in SEQ ID 1-4. The recombinant protein MBD-TET formed by fusing the MBD and the TET enzyme can be used for remarkably enhancing the oxidation activity of the TET enzyme, including NgTET1, mTET1CD, mTET2CDT, hTET1CD and hTET2CDT, on the 5mC. In addition, the invention further provides a simple, convenient and rapid high-throughput DNA methylation detection process, and the sensitivity and accuracy of a TET enzymatic oxidation reaction-based DNA methylation detection technology are improved. 本发明提供了增强TET酶活性的重组蛋白结构域，为DNA甲基化结合结构域（MBD），MBD1-4的氨基酸序列如SEQ ID 1-4所示。通过MBD与TET酶融合形成的重组蛋白MBD-TET可以显著增强TET酶包括NgTET1、mTET1CD、mTET2CDT、hTET1CD和hTET2CDT对5mC的氧化活性。此外，本发明还提供了简便快速的高通量DNA甲基化检测流程，提高基于TET酶氧化反应的DNA甲基化检测技术的灵敏度和准确性。