2-haloacid dehalogenase plus laccase with promoter and ribosome binding site
This part is the promoter circuit of BBa_K2546002. We add a promoter which is come from BBa_J23100 in front of the HAD+Laccase to let it express. Thus we can test the growth curve with TCDD and without TCDD.
First, we transform the BBa_J23100 into E.coli(DH5alpha), to let it amplify the plasmid.
Then we cut BBa_K2546002 with XbaI and PstI, and cut the BBa_J23100 with SpeI and PstI. After ligation and transformation, we got the colony and do the liquid LB incubate in the 37℃ overnight. In the next day, we extract the plasmid and get the result below.
The ideal plasmid is 4560 bp , and the result is match. We also operate PCR to check.
The band is nearly below the 3000bp, which is similar to 2756bp. Eventually, we successful assemble the HL composite part.
the Growth Curve Test
The growing of HAD+TLcc (J23100) got just slightly attenuate by TCDD (Fig. 1B) contrast to the growing of the control J23100 (Fig. 1A).
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 7
Illegal NheI site found at 30
- 21Illegal BglII site found at 745
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 394
Illegal NgoMIV site found at 491
Illegal NgoMIV site found at 1216
Illegal AgeI site found at 734
Illegal AgeI site found at 1726
Illegal AgeI site found at 2110
- 1000COMPATIBLE WITH RFC