Genetic Transformation by Heat Shock
The Effects of Using the Heat Shock Treatment to Deliver a Vector in Genetic Transformation
Introduction:
In this experiment we are testing what is required for E. coli to successfully grow on LB (Luria Broth) plates with ampicillin and determining if any genetic transformation has occurred. By combining +pGLO LB and ampicillin we should get an ampicillin resistant gene and by using –pGLO we should create a non-genetic resistant bacteria. The pGLO plasmid has the GFP (green fluorescent protein) gene and the gene that allows the plasmid to be resistant to the antibiotic ampicillin. The most important part of this experiment is the “heat shock treatment” because the E. coli membrane becomes permeable and increases the competency of the cells which in the end enhances the rate of successful transfer. According to Weedman (2009) genetic transformation is using foreign DNA and inserting it into an organism is known as “change in genes.” This is common in biotechnology in the agricultural sector, bioremediation, and in the field of medicine. In agricultural sector, genes can be genetically changed into plant species that originally lacked that trait. In bioremediation, a bacterium is altered with genes that allow them to digest oil spills. Furthermore in medicine, diseases can be treated with healthy copies of the defective genes which could save the individual. Weedman (2009) also states three different ways that genetic transformation can occur. Those include projective bombardment, electroporation, and heat shock. In projectile bombardment cells are attacked with tungsten pellets coated with foreign strands of DNA with a gene gun. In electroporation cells are bathed in a liquid that contains foreign DNA and is given a series of electrical pulses which increases the permeability of the cell membrane and causes the cells to take up the DNA from the surrounding medium. Heat shock does pretty much the same thing as electroporation except that the cells are
Introduction:
In this experiment we are testing what is required for E. coli to successfully grow on LB (Luria Broth) plates with ampicillin and determining if any genetic transformation has occurred. By combining +pGLO LB and ampicillin we should get an ampicillin resistant gene and by using –pGLO we should create a non-genetic resistant bacteria. The pGLO plasmid has the GFP (green fluorescent protein) gene and the gene that allows the plasmid to be resistant to the antibiotic ampicillin. The most important part of this experiment is the “heat shock treatment” because the E. coli membrane becomes permeable and increases the competency of the cells which in the end enhances the rate of successful transfer. According to Weedman (2009) genetic transformation is using foreign DNA and inserting it into an organism is known as “change in genes.” This is common in biotechnology in the agricultural sector, bioremediation, and in the field of medicine. In agricultural sector, genes can be genetically changed into plant species that originally lacked that trait. In bioremediation, a bacterium is altered with genes that allow them to digest oil spills. Furthermore in medicine, diseases can be treated with healthy copies of the defective genes which could save the individual. Weedman (2009) also states three different ways that genetic transformation can occur. Those include projective bombardment, electroporation, and heat shock. In projectile bombardment cells are attacked with tungsten pellets coated with foreign strands of DNA with a gene gun. In electroporation cells are bathed in a liquid that contains foreign DNA and is given a series of electrical pulses which increases the permeability of the cell membrane and causes the cells to take up the DNA from the surrounding medium. Heat shock does pretty much the same thing as electroporation except that the cells are
Cited: Kikkert, R. J. "Biological Projectiles (phage, Yeast, Bacteria) for Genetic Transformation of Plants." REFDOC. Web. 04 Nov. 2010. <http://cat.inist.fr/?aModele=afficheN&cpsidt=1703156>. Zhang, G. "Enhancing Electro-transformation Competency of Recalcitrant Bacillus Amyloliquefaciens by Combining Cell-wall Weakening and Cell-membrane Fluidity Disturbing." Analytical Biochemistry (2010). Print. Zhou, L. "Long-term Protection against Human Papillomavirus E7-positive Tumor by a Single Vaccination of Adeno-associated Virus Vectors Encoding a Fusion Protein of Inactivated E7 of Human Papillomavirus 16/18 and Heat Shock Protein 70." Human Gene Therapy 21.1 (2010): 109-19. Print.