Difference between revisions of "Part:BBa K4385017"
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This part consists of the constitutive promoters J23100, OmpC (BBa_K4385003) and ZBP (BBa_K4385004), which is used to verify the function of the zinc ion surface display parts. The 6*His tag was added to this sequence to purify the recombinant protein. | This part consists of the constitutive promoters J23100, OmpC (BBa_K4385003) and ZBP (BBa_K4385004), which is used to verify the function of the zinc ion surface display parts. The 6*His tag was added to this sequence to purify the recombinant protein. | ||
− | + | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | OmpC-ZBP can be anchored on the surface of E. coli cell membrane and enhance its absorption capacity of zinc. | ||
+ | ===Characterization=== | ||
+ | The constitutive promoter PJ23100 was placed upstream of OmpC-ZBP to verify its function (Fig. 1A). To verify the construction of PJ23100-OmpC-ZBP-ST, the enzyme digestion and gel electrophoresis were performed (Fig. 1B). | ||
+ | The concentration of zinc was measured by Micro Serum Zinc Concentration Assay Kit. As shown in Fig. 1C, the concentration of zinc ions in the supernatant is lower than that of the control. And the absorption capacity of cells is higher than that in the control, which indicate that our engineered bacteria can better absorb zinc ions. | ||
+ | [[Image: Zn_csd.png|center|frame|100px|<b>Figure 1.Validation of the absorption of Zn2+.</b>(A) Digestion and gel electrophoresis of PZraP-OmpC-ZBP-ST (B) The measurement of the concentration of Zn2+ in the supernatant. The Zn2+ was added when the OD600 value of bacteria reached 0.5. The concentration in supernatant was measured after 4 hrs of induction and normalized by the same OD600 value. *,P<0.05 from control group. △,P<0.05 from respective control.]]<br><br> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 03:55, 12 October 2022
Zinc constitutive surface display
This part consists of the constitutive promoters J23100, OmpC (BBa_K4385003) and ZBP (BBa_K4385004), which is used to verify the function of the zinc ion surface display parts. The 6*His tag was added to this sequence to purify the recombinant protein.
Usage and Biology
OmpC-ZBP can be anchored on the surface of E. coli cell membrane and enhance its absorption capacity of zinc.
Characterization
The constitutive promoter PJ23100 was placed upstream of OmpC-ZBP to verify its function (Fig. 1A). To verify the construction of PJ23100-OmpC-ZBP-ST, the enzyme digestion and gel electrophoresis were performed (Fig. 1B). The concentration of zinc was measured by Micro Serum Zinc Concentration Assay Kit. As shown in Fig. 1C, the concentration of zinc ions in the supernatant is lower than that of the control. And the absorption capacity of cells is higher than that in the control, which indicate that our engineered bacteria can better absorb zinc ions.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 414
Illegal SpeI site found at 573
Illegal PstI site found at 921 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 414
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 1304
Illegal NheI site found at 1327
Illegal SpeI site found at 573
Illegal PstI site found at 921 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 414
Illegal BglII site found at 122
Illegal BglII site found at 1439 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 414
Illegal SpeI site found at 573
Illegal PstI site found at 921 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 414
Illegal SpeI site found at 573
Illegal PstI site found at 921
Illegal AgeI site found at 240 - 1000COMPATIBLE WITH RFC[1000]