In addition, diseases such as Huntington’s disease, breast cancer, and muscular dystrophy are presently being screened for in humans (Jaroff, 1996). How researchers are able to screen for genes New developments have given researchers the ability to decipher the genetic code of organisms. Some of the techniques that researchers use are RFLP (restriction fragment length polymorphism) analysis and DNA probes. RFLP analysis utilizes enzymes from bacteria that are thought to be used as defense mechanisms against invading viral DNA. The enzymes fragment foreign DNA at specific locations depending on the base sequence (Griffiths, 1996).
If an antigen is identified as a disease causing agent, a monoclonal antibody could be an appropriate therapeutic agent to develop. Or if a single protein is likely to exert a pharmacological effect recombinant DNA may be appropriate. After the desired chemical substances have been identified, research for production can begin. The processes of recombinant DNA production and Monoclonal antibody production are very different. Most antigens have many antibody binding sites, or epitopes.
With this they were able to find that their protein was dispersed throughout the cytoskeletal structure of the leukocytes. After finding them they treated they with cytochalasin D to then examine the increases in distribution and... ... middle of paper ... ...unction as to tag any sort of particular protein. Understanding how antibodies interact with proteins and other microscopic structures also takes a comprehension of cytology, or the structure of these antibodies and their specificity for structures. These antibodies function similarly on structure as a lock-and-key mechanism with a specific antibody for a specific protein. Finally for any of this to function on paper and in the lab there must be genetic coding and expression of this code for these proteins and these antibodies to be produced.
The Use of Recombinant DNA Technology Recombinant DNA technology is the technology of preparing recombinant DNA in vitro by cutting up DNA molecules and splicing together fragments from more than one organism. (1) This is the process of using recombinant DNA technology to enable the rapid production of human protein from a single gene of insulin. Firstly the single gene required must be isolated. This can be done three ways: Either by working backwards from the protein- Finding the amino acid sequence for the protein needed, the order of bases can be established using known genetic code. New DNA can be made from this sequence of bases resulting in artificial gene made from complementary DNA.
The antigen binding site is constructed from VL and VH domains of the antibody molecule whereby sequence in this region is highly variable (Watson et al., 2008)). There is also domain of the antibody where the regions do not differ among different antibody molecules and is called “C” or constant. In developing B cells, DNA sequences of immunoglobulin unable to express directly from germ line so the individual gene segments must be rearrange to assemble a functional gene. During the development of B cells the V and J light-chain segments are spliced and join random by somatic recombination process. These segments are then brought together with CL-coding region by RNA splicing.
There are several applications for genetic engineering in microbiology as well as other fields of biology.It includes invitro mutagenesis,gene synthesis, Expressing eukaryotic genes in bacteria,production of transgenic plants and animals,gene therapy,screening for genetic diseases and forensic analysis. STEPS IN RECOMBINANT DNA TECHNOLOGY 1) PURIFYING DNA/ISOLATION OF DNA Purifying DNA and isolating genes is an important step in geneticengine... ... middle of paper ... ...e for the synthesis of new DNA strands, each randomly terminating due to the incorporation of a chain terminating dideoxynucleotide in 4 different reaction tubes. This produces a population of molecules, each terminating at a different site. Running the products in each tube on a gel allows the determination of where each chain terminating dideoxynucleotide was incorporated. The DNA is visualized because the DNA primer to start the reaction is radioactive or some of the dNTPs are radioactive This procedure is now automated so that a computer reads the sequence.
The molecules will then travel through the gel in different directions and speeds, based on their size and charge, allowing them to be separated from each other. Dyes, fluorescent tags, and radioactive labels can all enable the molecules on the gel to be seen after they have been separated. Because of these identification markers, they appear as a band across the top of the gel. Electrophoresis can be used for many different things. It is used to identify and study DNA or DNA fragments, and helps us to better understand the molecular components of both living and deceased organisms.
The purpose of DNA extraction is to obtain DNA in a relatively purified form which can be used for further investigations such as PCR and DNA sequencing. Many different methods are available for isolating genomic DNA. Modification and optimization of DNA extraction methods are usually done for different types of cell (Dhaliwal, 2013). Cetyltrimethyl ammonium bromide (CTAB) is a preferable method that can be used for isolating the genomic DNA. There are four important steps in DNA extraction protocol of animal tissue which are enzymatic digestion of cellular protein, DNA precipitation, DNA washing, and DNA hydration (refer to Figure 1 in Appendix 1).
Genetic engineering is possible because of special enzymes that cut DNA. These enzymes are called restriction enzymes or restriction endonucleases. Restriction enzymes are proteins produced by bacteria to prevent or restrict invasion by foreign DNA. They act as DNA scissors, cutting the foreign DNA into pieces so that it cannot function. A nuclease is any enzyme that cuts the phosphodiester bonds of the DNA backbone, and an endonuclease is an enzyme that cuts somewhere within a DNA molecule.
They then release enzymes to break down the substance. In the experiment, c opper acted as the foreign body while QM212 was the antibody. One application of this process could be used by the military. The military, utilizing bio-chemical tools, could engineer an antibody which binds with nerve gas and splits each molecule. This could be accomplished by first of all searching t he Brookhaven database for a proper antibody.