Difference between revisions of "Part:BBa K1806006"
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The natural urease of the bacteria specie S. pasteruii | The natural urease of the bacteria specie S. pasteruii | ||
Urease catalyses the breakdown of urea to ammonia. This breakdown is a major endothermic process, conducting a high efficiency endothermic process. | Urease catalyses the breakdown of urea to ammonia. This breakdown is a major endothermic process, conducting a high efficiency endothermic process. | ||
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+ | == Cloning == | ||
+ | |||
+ | The sponsors providing the parts were not able to synthesize orders of more than 2000 bp. For this reason, the part was split into two respective segment parts to be ligated. The two segments were tagged as G-Block 10 and 11. | ||
+ | Initially the G-Blocks were ligated to the pSB1C3 vector to be cloned. The verified cloning of the segment parts can be seen in the gel images below. | ||
+ | |||
+ | [[File:6006 -1.png]] [[File:6006-2.png]] | ||
+ | |||
+ | The two segment parts were ligated from the Hind3 restriction site. The ligated 10+11 urease complete part was cut with the EcoR1 and Pst1 restriction enzymes. The bands had the appropriate base lengths in the gel runs. | ||
+ | |||
+ | [[File:6006-3.png]] | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 01:46, 20 September 2015
Urease S. pasteruii
The natural urease of the bacteria specie S. pasteruii Urease catalyses the breakdown of urea to ammonia. This breakdown is a major endothermic process, conducting a high efficiency endothermic process.
Cloning
The sponsors providing the parts were not able to synthesize orders of more than 2000 bp. For this reason, the part was split into two respective segment parts to be ligated. The two segments were tagged as G-Block 10 and 11. Initially the G-Blocks were ligated to the pSB1C3 vector to be cloned. The verified cloning of the segment parts can be seen in the gel images below.
The two segment parts were ligated from the Hind3 restriction site. The ligated 10+11 urease complete part was cut with the EcoR1 and Pst1 restriction enzymes. The bands had the appropriate base lengths in the gel runs.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 206
Illegal BamHI site found at 2
Illegal XhoI site found at 2506 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 494
Illegal NgoMIV site found at 876
Illegal AgeI site found at 1673 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1629