Difference between revisions of "Part:BBa K732002"
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<partinfo>BBa_K732002 short</partinfo> | <partinfo>BBa_K732002 short</partinfo> | ||
This construct contains a strong constitutive promoter (J23119) as well as an RBS (B0034) upstream of synCG-β (K732001), and should strongly express a synthetic version of the beta subunit of human chorionic gonadotropin. | This construct contains a strong constitutive promoter (J23119) as well as an RBS (B0034) upstream of synCG-β (K732001), and should strongly express a synthetic version of the beta subunit of human chorionic gonadotropin. | ||
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| + | One application of this component is detection by pregnancy test. | ||
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| + | '''Proof-of-principle''' | ||
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| + | When heterogeneous mixture of ''E. coli'' expressing BBa_K732001 and BBa_K732002 was lysed by either sonication or T7 infection, re-folded using a literature-based protocol, and applied to NewChoice pregnancy tests (purchased from Dollar Tree), positive results were visible. | ||
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| + | Note that (a) there was some coloration around the positive band, and (b) the pregnancy tests were left overnight for bands to become visible. We hypothesize that the delayed read-time is necessary because the folding solution interferes with band visibility. | ||
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| + | [[Image:Virginia 2012 Oct_2_Preg_Test_Results.jpg]] | ||
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Revision as of 04:29, 10 October 2012
synCG-β + Promoter + RBS
This construct contains a strong constitutive promoter (J23119) as well as an RBS (B0034) upstream of synCG-β (K732001), and should strongly express a synthetic version of the beta subunit of human chorionic gonadotropin.
One application of this component is detection by pregnancy test.
Proof-of-principle
When heterogeneous mixture of E. coli expressing BBa_K732001 and BBa_K732002 was lysed by either sonication or T7 infection, re-folded using a literature-based protocol, and applied to NewChoice pregnancy tests (purchased from Dollar Tree), positive results were visible.
Note that (a) there was some coloration around the positive band, and (b) the pregnancy tests were left overnight for bands to become visible. We hypothesize that the delayed read-time is necessary because the folding solution interferes with band visibility.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 224
Illegal NgoMIV site found at 268 - 1000COMPATIBLE WITH RFC[1000]