重组DNA技术Recombinant DNA Technology.pdfVIP

Recombinant DNA Technology Key Lab of Molecular Medicine, Ministry of Education Duan Ma, Prof, MD, PhD Recombinant DNA 重组ＤＮＡ  Recombinant DNA is a form of synthetic DNA that is engineered through the combination or insertion of one or more DNA strands.  Through the use of recombinant DNA, genes that are identified as important can be amplified and isolated for use in other species or applications, where there may be some form of genetic illness. Some Definition  Clone: A group of organisms or cells produced asexually from one ancestor or stock, to which they are genetically identical.  Cloning: Propagation of a DNA sequence by incorporating it into a hybrid construct that can be replicated in a host cell.  Reproductive cloning: Process of making a genetically identical copy of an organism.  Therapeutic cloning: Process of making multiple copies of a cell to treat a disease. What is Cloned DNA Used For?  Work out the function of a gene.  Investigate a gene’s characteristics (size, expression, tissue distribution).  Look at how mutations may affect a gene’s function.  Make large concentrations of the protein coded for by the gene. Main Steps in DNA Cloning  The chosen piece of DNA is ‘cut’ from the source organism using restriction enzymes.  The piece of DNA is ‘pasted’ into a vector and the ends of the DNA are joined with the vector DNA by ligation.  The vector is introduced into a host cell, by a process called transformation. The host cells copy the vector DNA along with their own DNA, creating multiple copies of the inserted DNA.  The vector DNA is isolated from the host cells’ DNA and purified. How rInsulin Produced? Molecular Tools in Recombinant DNA Technology Restriction Endonuclease