Most useful to the fields of biochemistry and molecular genetics is the use of these methods in gene identification. First a gene must be isolated from an organism. This can be accomplished using restriction enzymes, cutting the DNA into pieces and then inserting these pieces into plasmid vectors, creating a library of genes. These vectors are then inserted into bacteria, which proceed in replicating the genes and producing their products. Any bacteria producing the protein of interest are isolated, using radiolabeled antibodies which bind specifically to the target protein.
These segments are then brought together with CL-coding region by RNA splicing. Recombination produces variants of antibody chain. There are several mechanisms of generation of antibody diversity in human. Figure 1 below shows the f... ... middle of paper ... ...r discarded (Watson et al., 2008). Works Cited Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., & Walter, P. (2002).
Toll-like receptors are proteins that play a significant role in the recognition of pathogens such as viruses and bacteria, thereby initiating an innate response. The TLR family recognizes structurally conserve molecules known as pathogen-associated molecular patterns (PAMPS) derive from these viruses and bacteria. TLR7 have been shown to mediate the recognition of single stranded RNA (ssRNA) viruses, whereas TLR9 recognizes non-methylated CpG sequences in bacterial DNA molecules. However, the molecular mechanism by which non-methylated CpG DNA and ssRNA viruses activate the innate immunity is not well understood. In this paper, it is demonstrated that the cell response to vesicular stomatitis viruses (VSV) and bacteria DNA is mediated by TLR7 and TLR9.
Scientists have tried to take advantage of this capability and manipulate the virus genome to remove disease-causing genes and insert therapeutic genes. Target cells such as the patient's liver or lung cells are infected with the viral vector. The vector then unloads its genetic material containing the therapeutic human gene into the target cell. The generation of a functional protein product from the therapeutic gene restores the target cell to a normal state. See a diagram depicting this process.
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).
1. (a) I. Plasmids are important tools in molecular biology. Plasmids are small circular DNA that has the ability to enter and replicate in bacterial cells and can be used as vectors to introduce foreign genes into bacteria for cloning and sequencing. Any gene must be inserted into an appropriate location of a plasmid to be expressed. The importance of a plasmid is in the step of cloning and sequencing when the construction of a recombinant DNA molecule occurs.
However under some conditions such as exposition of the bacterial cells to chemicals or to ultraviolet light, the phage DNA detaches from the bacterial DNA and enter a lytic cycle (Chauthaiwale 579). In the current study, the Southern blot technique was applied to identify the location of a DNA fragment within the lambda genome, which is also contained in a recombinant plasmid. Two restriction enzymes were used for the digestion of the DNA and the plasmid and a biotinylated probe was used for the hybridization.
Recombinant DNA technology and gene cloning have been fundamental to our understanding of gene structure and function. Recombinant DNA refers to the creation of new combinations of DNA segments that are not found together in nature. The isolation and manipulation of genes allows for more precise genetic analysis as well as practical applications in medicine, agriculture, and industry. An overview of recombinant dna technology is as follows: Isolate DNA /purifying DNA Cut with restriction enzymes Ligate into cloning vector transform recombinant DNA molecule into host cell each transformed cell will divide many times to form a colony of millions of cells, each of which carries the recombinant DNA molecule (DNA clone). 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.
They are antibodies that are mass produced in a laboratory from a single clone that recognizes only one epitope of a single antigen. Recombinant DNA is DNA that has been created artificially and codes for a specific desired protein for harvest. New biotechnology drug discovery begins with identification of cellular and genetic factors that play a role in specific diseases. Chemical and biological substances that target these factors are likely to have drug-like effects and are searched for. If an antigen is identified as a disease causing agent, a monoclonal antibody could be an appropriate therapeutic agent to develop.
The basic principles of EST is messenger RNA (mRNA) sequences to represent duplicates from expressed genes, cDNA libraries created from various cells and tissues, single pass sequencing of both ends of insert and accumulation of readable sequence part in EST database. In order to generate EST, RNA are first reverse transcr... ... middle of paper ... ...gs in Bioinformatics, 8(1), 6-21. Patino, W. D., Milan, O. Y., & Hwang, P. M. (2002). Serial analysis of gene expression. Circulation Research, 91, 565-569.