Recombinant T4 ligase mutant, encoded DNA and NGS library building method

The invention relates to a recombinant T4 ligase mutant, an encoded DNA and an NGS library building method. On the basis of wild type T4DL, Q19K, L63T, E88R, P127K, K159S, K225A, F233A, A237R, D371W, E440K, T451K and D452P multiple point mutation is performed; two double-stranded DNA binding domains...

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Bibliographische Detailangaben
Hauptverfasser:	LIU QIAN, SONG DONGLIANG, CAO ZHEN, HOU CE, CHEN JINGJING, WANG MAN, SUN RUI, 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 PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Beschreibung
Zusammenfassung:	The invention relates to a recombinant T4 ligase mutant, an encoded DNA and an NGS library building method. On the basis of wild type T4DL, Q19K, L63T, E88R, P127K, K159S, K225A, F233A, A237R, D371W, E440K, T451K and D452P multiple point mutation is performed; two double-stranded DNA binding domains are contained at the two ends of the mutant; and a polypeptide bridge is used for connection between the double-stranded DNA binding domains and the T4DL. Finally, a series of low-preference and high-efficiency recombinant DNA ligase mutants T4DLm are obtained. The invention further discloses the encoded DNA and the NGS library building method. The T4DLm has remarkable advantages on blunt end connection, the connection efficiency is as high as 95%, and the preference is extremely low. An efficient, simple and convenient novel NGS library building technology is developed by utilizing the T4DLm, has the remarkable advantages of short consumed time, higher library yield, better uniformity, lower library self-connecti