Difference between revisions of "Part:BBa K4407010"

Latest revision as of 08:32, 8 October 2022

Pthl-dps，Expression of dps with Pthl promoter

This part is responsible for expressing the dps protein under the control of a Pthl promotor, a ribosome binding site (RBS) and a terminator. It is composed ofBBa_K3443002 (Pthl), BBa_K103015 (RBS), BBa_K4407001 (dps), and BBa_K3585002 (terminator). dps protects the bacteria from oxidative damage. It protects DNA against oxidative damage by binding with Fe2+ to prevent it from reacting with H2O2 (Fenton’s reaction). In our project, we used this device to enhance the viability of C. tyrobutyricum in aerobic environment.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Results

Plasmid construction

We amplified a plasmid pMTL-Pthl-BS2 offered by team NJTech_China to obtain a linearized pMTL-Pthl vector. Then, we obtained the fragment of dps gene from the genome of D. wulumuqiensis R12 by PCR. The dps gene fragment and linearized pMTL-Pthl vector were ligated by the Gibson assembly method. Then, we ran a colony PCR for the transformed bacterial colony (E .coli JM109), inoculated the positive bacterial colonies, and extracted the plasmids, which were then confirmed to be pMTL-Pthl-dps by gene sequencing.

Conjugation into C. tyrobutyricum

After the recombinant plasmid pMTL-Pthl-dps was conjugated into C. tyrobutyricum using E. coli CA434 as the donor strain. The expression of dps protein in the recombinant bacteria was confirmed by SDS-PAGE.

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 <partinfo>BBa_K4407010 short</partinfo>
-This part is responsible for expressing the dps protein under the control of a Pthl promotor, a ribosome binding site (RBS) and a terminator. It is composed of<partinfo> BBa_K3443002</partinfo> (Pthl), <partinfo>BBa_K103015</partinfo> (RBS), <partinfo>BBa_K4407001</partinfo> (dps), and BBa_K3585002 (terminator). dps protects the bacteria from oxidative damage. It protects DNA against oxidative damage by binding with Fe2+ to prevent it from reacting with H2O2 (Fenton’s reaction). In our project, we used this device to enhance the viability of C. tyrobutyricum in aerobic environment.
+This part is responsible for expressing the dps protein under the control of a Pthl promotor, a ribosome binding site (RBS) and a terminator. It is composed of<partinfo>BBa_K3443002</partinfo> (Pthl), <partinfo>BBa_K103015</partinfo> (RBS), <partinfo>BBa_K4407001</partinfo> (dps), and BBa_K3585002 (terminator). dps protects the bacteria from oxidative damage. It protects DNA against oxidative damage by binding with Fe2+ to prevent it from reacting with H2O2 (Fenton’s reaction). In our project, we used this device to enhance the viability of C. tyrobutyricum in aerobic environment.
 <!-- Add more about the biology of this part here