蛋白和酶工程摘要重点.ppt

What is biotech? 生物技术（biotechnology），有时也称生物工程，是指人们以现代生命科学为基础，结合其他基础科学的科学原理，采用先进的科学技术手段，按照预先的设计改造生物体或加工生物原料，为人类生产出所需产品或达到某种目的。生物技术是人们利用微生物、动植物体对物质原料进行加工，以提供产品来为社会服务的技术。 生物技术（biotechnology）也译成生物工程，生物学研究与应用的技术方面，包括，基因工程、细胞工程、发酵工程和酶工程，现代生物技术发展到高通量组学（omics）芯片技术、基因与基因组人工设计与合成生物学等系统生物技术。 1. amino acids and proteins 2. protein sources 3. protein purification 4. methods to determine protein structure 5. protein structure 6. protein folding Amino Acids Can Be Classified by R Group Titration Curves (滴定曲线) of AA Protein Therapeutics 1922 Insulin was first purified from bovine and porcine pancreas and used as daily injection Problems: the availability of animal pancreas; cost of insulin purification; immunological reactions. 1982 Solution: Isolation of human insulin gene; Engineering of E. coli to express human insulin by rDNA technology, growing vast quantities of bacteria, large-scale production of human insulin Result: Abundant and inexpensive human insulin with low immunogenicity and free from animal pancreatic substances. Where is it from? Automatic Peptide Synthesizer (ABI 433A) Natural sources are often rare and expensive Difficult to keep up with demand Hard to isolate product Lead to immune reactions (diff. species) Viral pathogen contamination Most applied proteins today are produced recombinantly Cheaper, safer, abundant supply Recombinant Sources Recombinant Methods Developed in 1970’s 1980’s Paul Berg (1973) restriction enzymes Herbert Boyer (1978) cloning human insulin into E. coli Two general approaches Expression in isolated cells Expression in transgenic plants/animals Cloning Process Gene of interest is cut out with restriction enzymes (RE) Host plasmid (circular chromosome) is cut with same REs Gene is inserted into plasmid and ligated with ligase New (engineered) plasmid inserted into bacterium (transform) Recombinant Protein Expression Systems Escherichia coli Other bacteria Pichia pastoris Other yeast Baculovirus Animal cell culture Plants Sheep/cows