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Usage of biotechnology with recombinant dna
Benefits and risks of recombinant dna
Usage of biotechnology with recombinant dna
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Recombinant DNA technology has opened the door for humans to isolate and purify virtually any known genomic sequence. The human genome is known to contain approximately 6x109 base pairs over a span of 23 pairs of chromosomes. Getting pure DNA samples from large genomes like ours is now made far easier thanks to recombinant DNA technology. In addition, functional regions can be investigated and studied in predetermined manners, giving us vital insight to the biochemical, molecular, and genetic properties of our DNA (Lodish et al., 2000). Recombinant DNA itself is any DNA molecule formed by joining DNA fragments from different sources. The most frequent manner that recombinant DNA is produced is by restriction digestion, followed by ligation of the complementary sticky ends via DNA ligase. Each step in the creation of the construct plays a vital role, and should not go unrecognized.
Plasmids are extrachromosomal DNA strands that usually found within bacteria, yeast, and even some eukaryotes. These DNA strands have the ability to self-replicate, just like chromosomal DNA within these ...
Once the recombinant plasmid was obtained, it was then inserted into E. coli cells through transformation. From a successful transformation, we expected the bacterial cells to translate the inserted EGFP sequence into its protein form. The bacteria cultures were plated on petri dishes containing growth supplement, Luria Broth (LB), an antibiotic: Kanamycin, and IPTG which induced the fluorescence property within successfully transformed bacterial colonies. Different variants of the petri dishes were also included as control and unknown.
The plasmids in lanes 3,4,8 and 9 have been digested using one restriction enzyme and had been cut at one restriction site, resulting in a linear molecule. Comparing lanes 3 and 4 to
The purpose of this experiment is to identify an unknown insert DNA by using plasmid DNA as a vector to duplicate the unknown insert DNA. The bacteria will then be transformed by having it take in the plasmid DNA, which will allow us to identify our unknown insert as either the cat gene or the kan gene.
Therefore colonies containing the non-recombinant pUC19 plasmid have a functional lacz’ gene appear blue on the agar and colonies containing recombinant pUC19 would have a non-functional lacz’ gene due to insertional inactivation and appear white on the growing medium.
The given DNA ladder sample and each individual ligation samples were run on 40ml of 0.8% agarose in 1x TAE buffer for approximately sixty minutes at 110V. The appropriate volume of 6x GelRed track dye was used after it was diluted to a final concentration of 1x and incubated for thirty minutes. Finally, the gel was illuminated under UV light and analyzed.
al. (1994) explain that a complementary DNA for GFP produces a fluorescent product when expressed in E. coli cells as the expression of GFP can be used to monitor gene expression and protein localization in living things. In this experiment, the heat shock method will be used to deliver a vector (plasmid) of GFP to transform and grow E. coli bacteria. Four plates containing Luria Bertani (LB) broth and either –pGLO or +pGLO will have E. coli bacteria added to it. The plate containing –pGLO (no pGLO) and LB will show growth as ampicillin will be present killing bacteria but no glowing because no arabinose will be present for glowing to be activated, the same result will be seen in the plate containing +pGLO, LB and ampicillin.
It has an outer membrane that contains lipopolysaccharides, a periplasmic space with a peptidoglycan layer, and an inner cytoplasmic membrane. It also consists of adhesive fimbriae. Some strains of E. coli are piliated and are capable of accepting, as well as transferring plasmid to and from other bacteria. This enables the bacteria under stressful or bad conditions to survive. Although its structure is simple with only one chromosomal DNA and a plasmid, it can perform complicated metabolism to help maintain its cell division and cell growth. E. coli produce very rapidly; a single microscopic cell can divide to form a visible colony with millions of cells overnight (phschool.com). It is the preferred bacteria in most laboratories because it grows fast and easy, and can obtain energy from a wide variety of sources. Since the birth of molecular cloning, E. coli has been used as a host for introduced DNA sequences (biotechlearn.org.nz). In 1973, Boyer and Cohen showed that two short pieces of DNA could be cut and pasted together, and returned to
In 1990, the first great stride of genetics took place. This was called the Human Genome Project, a large-scale operation that was designed to understand the human genome (genetic structure). Since its commencement, there have been many leaps and bounds that have taken place. For certain genetic issues that we once knew nothing about, we no...
In the past 40 years, scientists have developed and applied genetic engineering to alter the genetic make-up of organisms by manipulating their DNA. Scientists can use restriction enzymes to slice up a piece of DNA from an organism with the characteristics they want and spliced (joint) to a DNA from another organism. DNA that contains pieces from different species is called recombinant DNA, and it now has different genetic material from its original. When this DNA inserted back into the organism, it changes the organism’s trait. This technique is known as gene-splicing (Farndon 19).
Watson, J. D., Gilman, M., Witkowski, J., Zoller, M. (1992). Recombinant DNA. New York: W. H. Freeman and Company.
The birth of genetic engineering and recombinant DNA began in Stanford University, in the year 1970 (Hein). Biochemistry and medicine researchers were pursuing separate research pathways, yet these pathways converged to form what is now known as biotechnology (Hein). The biochemistry department was, at the time, focusing on an animal virus, and found a method of slicing DNA so cleanly that it would reform and go on to infect other cells. (Hein) The medical department focused on bacteria and developed a microscopic molecular messenger, that could not only carry a foreign “blueprint”, or message, but could also get the bacteria to read and copy the information. (Hein) One concept is needed to understand what happened at Stanford: how a bacterial “factory” turns “on” or “off”. (Hein) When a cell is dividing or producing a protein, it uses promoters (“on switches”) to start the process and terminators (“off switches”) to stop the process. (Hein) To form proteins, promoters and terminators are used to tell where the protein begins and where it ends. (Hein) In 1972 Herbert Boyer, a biochemist, provided Stanford with a bacterial enzyme called Eco R1. (Hein) This enzyme is used by bacteria to defend themselves against bacteriophages, or bacterial viruses. (Hein) The biochemistry department used this enzyme as a “molecular scalpel”, to cut a monkey virus called SV40. (Hein) What the Stanford researchers observed was that, when they did this, the virus reformed at the cleaved site in a circular manner. It later went on to infect other cells as if nothing had happened. (Hein) This proved that EcoR1 could cut the bonding sites on two different DNA strands, which could be combined using the “sticky ends” at the sites. (Hein). The contribution towards genetic engineering from the biochemistry department was the observations of EcoR1’s cleavage of
Discoveries in DNA, cell biology, evolution, and biotechnology have been among the major achievements in biology over the past 200 years with accelerated discoveries and insight’s over the last 50 years. Consider the progress we have made in these areas of human knowledge. Present at least three of the discoveries you find to be the most important and describe their significance to society, heath, and the culture of modern life.
The reconstruction of DNA has brought many cures against genetic diseases that before were undetectable. Although it is not a treatme...
Our DNA is very good at replicating itself without errors, but it’s also extraordinarily long — a book publishing an individual’s DNA would be roughly the length of 1000 copies of “War and Peace.” Due to DNA’s size, even a near perfect copying mechanism introduces some errors,
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.