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Lecture: Development and Application of a Highly Efficient and Fast Method for Multisplit Large Plasmid Construction

Posted: 2017-12-11

Time:December 11th, 2017, 10:00 am
Venue:Conference Room 406 in Biotechnology Building
Lecturer:LIU Zihe, Beijing University of Chemical Technology

Abstract:

It is a big challenge to obtain long and complex DNA constructs for systems construction in synthetic biology, where the DNA assembly method is the foundation in artificial systems design. However, the convention methods always only work in assembling small DNA with the length < 5 kb. Thus, novel efficient methods should be developed for synthesis of larger-scale DNA molecules.

The speaker's research team has developed a new DNA assembly technique-“Twin-primer non-enzymatic DNA assembly” (TPA). The TPA method relies mainly on the overlapping sequences of the fragments to be ligated, the fragments are bound to each other at a specific annealing temperature, and then introduced into E. coli to obtain the desired plasmid. Compared with the widely used Gibson assembly, the TPA method is superior to the Gibson method in the aspects of multi-fragment integration, large plasmid integration and high GC content fragments integration. The in vitro TPA DNA assembly technology, no requiring additional reagents, simplifies the procedure for routine genetic manipulation and saving reagents and time.

Introduction:

LIU Zihe, Ph.D., works as an Associate Professor in College of Life Science and Technology, Beijing University of Chemical Technology. She received her Ph.D. degree from Chalmers University of Technology, Sweden in 2012. From 2013 to 2017 worked as research scientist in Agency for Science, Technology and Research, Singapore. At present, she is employed in Beijing University of Chemical Technology engaged in the research of system biology and synthetic biology. Her research interests focus on the development of efficient, sustainable yeast cell factories that produce high value fuels and chemicals using synthetic biology tools, and the development of synthetic biology tools. Representative work was published in Nucleic Acids Research, ACS Synthetic Biology and with a total impact factor> 40.

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