You are interested in cloning a gene that codes for an enzyme that produces a blue pigment. You have chosen to use the plasmid pUC19 for cloning (shown below). Drdl 91 Acll 2297 Xmnl 2294 Bcgl 2215 Scal 2177 Pvul 2066 Avall 2059 BsrDI 1935 Acll 1924 Fspl 1919- Avall 1837 NmeAIII 1822 Bell 1813- Bpml 1784 BsrFI 1779 BciVI 2542 Aatll-Zral 2617 Sspl 2501 Bsal 1766 BsrDI 1753 Bmrl 1744 Ahdl 1694 BsmBI 51 BsmBI 2683 Eco01091 2674 PUC19 2,686 bp BoeAl 1292 ori BsTAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Bell 245 Fspl 256 lacZo AlwNI 1217 Pvul 276 Pvull 306 Bmrl 364 BseYI 1110 BceAl 387 MCS BeiVI 1015 Til 781 AflIII - Pcil 806 Drdl 908 Apol-EcoRI 396 Banll-Sacl - Eco53KI 402 Acc651- Kpnl 408 Aval - BsoBI - Smal TspMI - Xmal 412 Pvull 628 Tfil 641 BsaXI 659 BspQI-Sapl 683 BamHI 417 Xbal 423 Accl - Hincll-Sall 429 BspMI -BfuAI 433 Sbfl 434 Pstl 435 Sphl 441 Hind 447 The plasmid codes for three elements, lacZ, ApR and ori. What are these three things? Which restriction enzyme cut sites would you use to clone your blue pigment gene into pUC19 and why? You digest the plasmid and your blue pigment gene with the restriction enzymes chosen in step b. Explain how the two fragments will stick together, and explain how you will stich the phosphate backbone back up.
You are interested in cloning a gene that codes for an enzyme that produces a blue pigment. You have chosen to use the plasmid pUC19 for cloning (shown below). Drdl 91 Acll 2297 Xmnl 2294 Bcgl 2215 Scal 2177 Pvul 2066 Avall 2059 BsrDI 1935 Acll 1924 Fspl 1919- Avall 1837 NmeAIII 1822 Bell 1813- Bpml 1784 BsrFI 1779 BciVI 2542 Aatll-Zral 2617 Sspl 2501 Bsal 1766 BsrDI 1753 Bmrl 1744 Ahdl 1694 BsmBI 51 BsmBI 2683 Eco01091 2674 PUC19 2,686 bp BoeAl 1292 ori BsTAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Bell 245 Fspl 256 lacZo AlwNI 1217 Pvul 276 Pvull 306 Bmrl 364 BseYI 1110 BceAl 387 MCS BeiVI 1015 Til 781 AflIII - Pcil 806 Drdl 908 Apol-EcoRI 396 Banll-Sacl - Eco53KI 402 Acc651- Kpnl 408 Aval - BsoBI - Smal TspMI - Xmal 412 Pvull 628 Tfil 641 BsaXI 659 BspQI-Sapl 683 BamHI 417 Xbal 423 Accl - Hincll-Sall 429 BspMI -BfuAI 433 Sbfl 434 Pstl 435 Sphl 441 Hind 447 The plasmid codes for three elements, lacZ, ApR and ori. What are these three things? Which restriction enzyme cut sites would you use to clone your blue pigment gene into pUC19 and why? You digest the plasmid and your blue pigment gene with the restriction enzymes chosen in step b. Explain how the two fragments will stick together, and explain how you will stich the phosphate backbone back up.
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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Transcribed Image Text:You are interested in cloning a gene that codes for an enzyme that produces a blue
pigment. You have chosen to use the plasmid pUC19 for cloning (shown below).
Drdl 91
Acll 2297
Xmnl 2294
Bcgl 2215
Scal 2177
Pvul 2066
Avall 2059
BsrDI 1935
Acll 1924
Fspl 1919
Avall 1837
NmeAIII 1822
Bgll 1813-
Bpml 1784-
BsrFI 1779
Bsal 17661
BsrDI 1753
Bmrl 1744
Aatll - Zral 2617
BriVI 2542
Sspl 2501-
BsMBI 51
BsmBI 2683
Eco01091 2674
Ahdl 1694
PUC19
2,686 bp
BceAl 1292
ori
BstAPI 179
Ndel 183
Kasl - Narl - Sfol 235
Bell 245
Fspl 256
laczo
AlwNI 1217
Pvul 276
Pvull 306
Bmrl 364
BseYI 1110
BceAl 387
MCS
BeiVI 1015
Til 781
AflIlI-Pcil 806
Drdl 908
Apol-EcoRI 396
Banll - Sacl - Eco53KI 402
Acc651 - Kpnl 408
Aval-BsoBI- Smal-
TspMI-Xmal 412
Bamil 417
Pvull 628
Tfil 641
BsaXI 659
BspQI-Sapl 683
Xbal 423
Accl - Hincll - Sall 429
BspMI-BfuAl 433
Sbfl 434
Pstl 435
Sphl 441
HindIII 447
The plasmid codes for three elements, lacZ, ApR and ori. What are these three
things?
Which restriction enzyme cut sites would you use to clone your blue pigment gene
into pUC19 and why?
You digest the plasmid and your blue pigment gene with the restriction enzymes
chosen in step b. Explain how the two fragments will stick together, and explain
how you will stich the phosphate backbone back up.
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