基因合成及工业酶分子改造中的应用.ppt

Proceedings of the Gene Synthesis and application in enzyme genetic modification基因合成及工业酶分子改造中的应用 by YANG, JIANGKE 杨江科 School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University 武汉工业学院 WHAT IS SYNTHETIC BIOLOGY?什么是合成生物学 Synthesis: The combination of separate elements to form a coherent whole Synthetic biology: The design and construction of new biological parts, devices, and systems The re-design of existing, natural biological systems for useful purposes. GENE SYNTHESIS:THE METHOD AND STRATEGY基因合成的方法和策略 GENE SYNTHESIS STRATEGY Ligation - incredibly tedious and inefficient Dual asymmetic PCR Assembly PCR Two-step PCR A yeast alanine transfer RNA (77 bp) (a) The chemical synthesis of 15 polydeoxynueleotide segments ranging in length from 5 to 20 nucleotide units (b) Ligase-catalyzed covalent joining of several segments to form three parts of the duplex, (c) Joining of the three parts to construct the entire duplex. What is the next ? Now we have 14 “perfect” 750-mers We need to assemble the 750-mers into one 10 kb piece We need them to go together in exactly the right order and orientation We need an “Assembly reaction” A typical “palindromic overhang” is generated by most restriction enzymes View of the assembly reaction And extending the principles of design and construction to whole organisms Applications of Synthetic Biology合成生物学的应用 Biological workspace Microbes Mammalian cells Plants .... Broader applications Therapeutics (Pharmaceuticals, Biologics) Energy (esp. Fuels) Chemicals Agriculture …. Achievements in Synthetic Biology Therapeutics Achievements in Synthetic Biology Fuels Achievements in Synthetic Biology Fuels The work we have done and what we are doing我们所做的工作 Aims: Enzyme immobilization酶固定化 Protein Expression Protein/Structure Independent Factors: promoters and upstream elements translational initiation and termination mRNA stability codon bias Protein/Structure Dependent Factors: folding and aggregation pr