A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 Bsnlil 2683 Eco01091 2674 BstAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617. Bell 245 Vpl 256 Prul 276 Pull 306 BiVI 2542. Sspl 2501 Hmrl 364 Aell 2297 Bcetl 387 Apel - Ecoti 396 Banll - Sacl - EcosKI 402 lecs1 - Kpnl 408 Xmnl 2294 laczu Begl 2215 MCS wal - BoBI - Smal- Scal 2177 TspMI - Xmal 412 Hamill 417 thal 23 Prul 2066 Aecl - linell - Sall 429 Avall 2059 PUC19 2,686 bp Sh 434 Pstl 35 BaDI 1935 Acll 1924 Fipl 1919 Sphl 441 Hindl 417 Prull 628 wall 1837 NmeI 1822 Bell 1813 Bpml 1784 Bsrfl 1779 Bsal 1766 Til 641 BsaXI 659 Bspol - Sapl 683 Thl 781 BarD 1753 Bnrl 1744 ANIII - Peil 006 Drdl 908 ori Ahdl 1694 BeiVI 1015 Bsell 1110 AlwNI 1217 kell 1292 A. Which plasmid is used? B. Which antibiotic was used?
A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 Bsnlil 2683 Eco01091 2674 BstAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617. Bell 245 Vpl 256 Prul 276 Pull 306 BiVI 2542. Sspl 2501 Hmrl 364 Aell 2297 Bcetl 387 Apel - Ecoti 396 Banll - Sacl - EcosKI 402 lecs1 - Kpnl 408 Xmnl 2294 laczu Begl 2215 MCS wal - BoBI - Smal- Scal 2177 TspMI - Xmal 412 Hamill 417 thal 23 Prul 2066 Aecl - linell - Sall 429 Avall 2059 PUC19 2,686 bp Sh 434 Pstl 35 BaDI 1935 Acll 1924 Fipl 1919 Sphl 441 Hindl 417 Prull 628 wall 1837 NmeI 1822 Bell 1813 Bpml 1784 Bsrfl 1779 Bsal 1766 Til 641 BsaXI 659 Bspol - Sapl 683 Thl 781 BarD 1753 Bnrl 1744 ANIII - Peil 006 Drdl 908 ori Ahdl 1694 BeiVI 1015 Bsell 1110 AlwNI 1217 kell 1292 A. Which plasmid is used? B. Which antibiotic was used?
Human Anatomy & Physiology (11th Edition)
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Chapter1: The Human Body: An Orientation
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Concept explainers
Bacterial Genomics
The study of the morphological, physiological, and evolutionary aspects of the bacterial genome is referred to as bacterial genomics. This subdisciplinary field aids in understanding how genes are assembled into genomes. Further, bacterial or microbial genomics has helped researchers in understanding the pathogenicity of bacteria and other microbes.
Transformation Experiment in Bacteria
In the discovery of genetic material, the experiment conducted by Frederick Griffith on Streptococcus pneumonia proved to be a stepping stone.
Plasmids and Vectors
The DNA molecule that exists in a circular shape and is smaller in size which is capable of its replication is called Plasmids. In other words, it is called extra-chromosomal plasmid DNA. Vectors are the molecule which is capable of carrying genetic material which can be transferred into another cell and further carry out replication and expression. Plasmids can act as vectors.
Question
Transcribed Image Text:A. Read the following section to review what you learned about plasmids, transformation and antibiotic
selection (which antibiotic?)
Plasmids and Recombinant DNA Technology
A plasmid is a circular piece of extra-chromosomal DNA (between 2000-10,000 bp) that is normally found in
bacteria. In nature, the plasmid will often contain a gene that encodes a protein that allows the bacteria to be
resistant to antibiotics, or a protein that allows the bacteria to cause an infection. Plasmids were discovered by
researchers in the late sixties, and it was quickly realized that they could be used to amplify a gene of interest to
create recombinant DNA. The gene of interest is inserted into the plasmid at restriction endonuclease sites. The
plasmid can then be used as a vector to shuttle DNA to a different host/bacteria.
There are many different kinds of plasmids commercially available. All of them contain:
1. a selectable marker (i.e., a gene that encodes for antibiotic resistance, color change, etc)
2. an origin of replication (which is used by the DNA making machinery in the bacteria as the starting
point to make a copy of the plasmid)
3. a multiple cloning site (MCS). The multiple cloning site has many restriction enzyme sites and is used to
insert the DNA of interest. The multiple cloning site is usually in the middle of a reporter gene like Lac
Z.
A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs)
Indl 91
BamBI 51
BsmBl 2683
ISTAPI 179
Eco01091 2674
Ndel 183
Kasl - Narl - Sfol 235
Aatll - Zral 2617
Bell 245
Fspl 256
Prul 276
Pvull 306
BeVI 2542.
Sspl 2501
Bmrl 364
Aell 2297
Boell 387
Apol - EcoRI 396
Hanll - Sacl - EcosSKI 402
Xmnl 2294
laczu
lec651 - Kpnl 408
Begl 2215
MCS
Awal - BaoRI - Smal -
Scal 2177
TspMI - Xmal 412
Bamll 417
Nhal 423
Prul 2066
vall 2059
lel. Hindll. Sall 429
PUC19
2,686 bp
Shf 434
BseI 1935
Pstl 435
Sphl 441
Adl 1924
Hindlll 417
Fspl 1919
Pvull 628
Avall 1837
NmeAlII 1822
Bell 1813
Til 641
BsaXI 659
Bpml 1784
Bsrll 1779
Bsal 1766
Bspol - Sapl 683
Til 781
AfAIII - Pcil 806
BsrDI 1753
Drdl 908
Bmrl 1744
ori
Ahdl 1694
BeiVI 1015
BseYl 1110
AlwNI 1217
BeAl 1292
A. Which plasmid is used?
B. Which antibiotic was used?
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