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Which of the delivery techniques mentioned in Table 2 can be used to best transfer recombinant plasmids into E. coli cells

Which of the delivery techniques mentioned in Table 2 can be used to best transfer recombinant plasmids into E. coli cells

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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Which of the delivery techniques mentioned in Table 2 can be used to best transfer recombinant plasmids into E. coli cells

COLUMBAN COLLEGE IN., OLONGAPO CITY SENIOR HIGH SCHOOL - BASIC EDUCATION DEPARTMENT ASINAN Name of microi into re LEARNER'S MODULE Name of the Student: Section: electric shock (zapping cells with electricity) is used. This technique involves wrapping the recombinant plasmids in vesicle-like bubbles which fuse with the recipient cells' membranes, thereby delivering the plasmids into the cells. In this technique, the recombinant plasmids are hitched or packed into viruses which are allowed to infect the recipient cells (usually gametes or zygotes of animals). Using Lipid Membrane Bubbles Using Viruses At this point, please turn to Assessment #1 and answer GQ#2. In order to identify which of the surviving recipient cells have successfully taken in the recombinant plasmid, scientists devised screening processes that will point out the transgenic
Transcribed Image Text:COLUMBAN COLLEGE IN., OLONGAPO CITY SENIOR HIGH SCHOOL - BASIC EDUCATION DEPARTMENT ASINAN Name of microi into re LEARNER'S MODULE Name of the Student: Section: electric shock (zapping cells with electricity) is used. This technique involves wrapping the recombinant plasmids in vesicle-like bubbles which fuse with the recipient cells' membranes, thereby delivering the plasmids into the cells. In this technique, the recombinant plasmids are hitched or packed into viruses which are allowed to infect the recipient cells (usually gametes or zygotes of animals). Using Lipid Membrane Bubbles Using Viruses At this point, please turn to Assessment #1 and answer GQ#2. In order to identify which of the surviving recipient cells have successfully taken in the recombinant plasmid, scientists devised screening processes that will point out the transgenic
ng the The next step involves the transfer of the recombinant plasmid into recipient cells. This may be done in several ways (depending on the nature of the recipient cells), some of which are briefly described in Table 2. It is important to note, however, that some of these delivery techniques may kill or destroy some of the recipient cells. When this happens, only the surviving cells become available for the succeeding screening process. Table 2: Techniques for Delivering Recombinant Plasmids into Recipient Cells Technique Description This technique uses gene guns to shoot or bombard plant cells with pellets which are coated with recombinant plasmids. In this technique, bacterial cells are first exposed to calcium chloride (CaCl2) in order to increase the pore sizes of their cell membranes. The cells are then incubated with the recombinant plasmid at about 4°C for around 30 minutes, allowing the plasmids to gather near the bacterial cells. The bacterial cell-plasmid solution is afterwards incubated at 42°C for 1 minute, then back to 4°C for 2 minutes. This heat shock (rapid increase and decrease of temperature) causes the cell membrane pores to increase and decrease in size, therefore allowing the recombinant plasmids near the membrane to enter the recipient cells. The procedure for this technique, which is commonly used for mammalian recipient cells, is very similar to heat shock treatment. The main difference is that, instead of using heat shock to increase the size of membrane pores, Biolistics Heat Shock Treatment Electroporation
Transcribed Image Text:ng the The next step involves the transfer of the recombinant plasmid into recipient cells. This may be done in several ways (depending on the nature of the recipient cells), some of which are briefly described in Table 2. It is important to note, however, that some of these delivery techniques may kill or destroy some of the recipient cells. When this happens, only the surviving cells become available for the succeeding screening process. Table 2: Techniques for Delivering Recombinant Plasmids into Recipient Cells Technique Description This technique uses gene guns to shoot or bombard plant cells with pellets which are coated with recombinant plasmids. In this technique, bacterial cells are first exposed to calcium chloride (CaCl2) in order to increase the pore sizes of their cell membranes. The cells are then incubated with the recombinant plasmid at about 4°C for around 30 minutes, allowing the plasmids to gather near the bacterial cells. The bacterial cell-plasmid solution is afterwards incubated at 42°C for 1 minute, then back to 4°C for 2 minutes. This heat shock (rapid increase and decrease of temperature) causes the cell membrane pores to increase and decrease in size, therefore allowing the recombinant plasmids near the membrane to enter the recipient cells. The procedure for this technique, which is commonly used for mammalian recipient cells, is very similar to heat shock treatment. The main difference is that, instead of using heat shock to increase the size of membrane pores, Biolistics Heat Shock Treatment Electroporation
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