Creating and Utilizing Recombinant DNA
First a sample of donor DNA is obtained and is usually the entire genome of a selected organism. Specific sections of the donor DNA may be obtained with the help of restriction enzymes. Restriction enzymes cut, or lyse, a section of DNA at a specific pre-determined nucleotide sequence. By selecting restriction enzymes at both ends of the desired DNA segment that specific sequence may be isolated. These segments of DNA can then be inserted into a vector, or the method of transport. Vectors are usually plasmids or modified bacteriophages. We have taken advantage of the natural mechanisms that these organisms use to reproduce by removing the genetic information that makes these vectors harmful and inserting desired information that we wish to be transferred. Complimentary strands of nucleotides can be utilized to insert desired information either into the normal location for this genetic segments or into a site where normal non-coding introns would be located. These vectors are then introduced into a cell type generally to provide for a protein deficiency (for a more specific example see the overview section on Bubble boy syndrome). This methodology can be applied for synthetically producing mass quantities of things that are normally unable to be produced synthetically. The polymerase chain reaction allows us to take advantage of recombinant DNA to produce large quantities of a desired protein in bacterial cells.
When using recombinant DNA in conjunction with gene therapy, the idea is to remove cells from a patient (in hopes that these cells wont illicit an immune response when reintroduced) and use a vector to insert genetic information to compensate for the hereditary defect intrinsic to that cell type. Germline therapy methods have been suggested which would involve altering the embryo through the use of stem cells and genetically recombined normal wild type function information. The technology for this has not been perfected and opens a number of ethical issues.
Resources:
Introduction to Genetic Analysis 8th edition by Griffiths, Wessler, Lewontin, Gelbart, Suzuki, Miller
This website was created for the course Honors 210:The Ideal at Monmouth College.