Difference between revisions of "Part:BBa K4248007"
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==Contribution== | ==Contribution== | ||
| − | LL-37 is expressed widely in human body cells, such as epithelial skin cell，bone marrow cells, and neutrophils. It is responsible for immune defense against bacterial invasion, with study showing its effect of significant reduction in attachment of bacteria and biofilm production | + | LL-37 is expressed widely in human body cells, such as epithelial skin cell，bone marrow cells, and neutrophils. It is responsible for immune defense against bacterial invasion, with study showing its effect of significant reduction in attachment of bacteria and biofilm production. |
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Because the T7 promoter and T7 RNA polymerase have strong ability in translation and usually be used as protein expression, we choose pET28a-vector and E.coli BL21(DE3), with T7 promoter and T7 RNA polymerase respectively, to express our target protein LL-37. To achieve this, we designed the DNA sequences of LL-37 to be inserted into the XbaI and XhoI sites of the pET-28a(+) vector (Figure 1.), transferred the recombinant plasmid into E.coli BL21(DE3) for protein expression. | Because the T7 promoter and T7 RNA polymerase have strong ability in translation and usually be used as protein expression, we choose pET28a-vector and E.coli BL21(DE3), with T7 promoter and T7 RNA polymerase respectively, to express our target protein LL-37. To achieve this, we designed the DNA sequences of LL-37 to be inserted into the XbaI and XhoI sites of the pET-28a(+) vector (Figure 1.), transferred the recombinant plasmid into E.coli BL21(DE3) for protein expression. | ||
[[File:T--Shanghai city-- BBa K4248007-figure1.png|500px|thumb|center|Figure 1. The map of recombinant plasmid pET-28a(+)- LL37.]] | [[File:T--Shanghai city-- BBa K4248007-figure1.png|500px|thumb|center|Figure 1. The map of recombinant plasmid pET-28a(+)- LL37.]] | ||
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Latest revision as of 09:49, 26 September 2022
T7 pro-6His - LL-37- T7 ter
T7 pro-6His - LL-37- T7 ter
Contribution
LL-37 is expressed widely in human body cells, such as epithelial skin cell，bone marrow cells, and neutrophils. It is responsible for immune defense against bacterial invasion, with study showing its effect of significant reduction in attachment of bacteria and biofilm production.
Because the T7 promoter and T7 RNA polymerase have strong ability in translation and usually be used as protein expression, we choose pET28a-vector and E.coli BL21(DE3), with T7 promoter and T7 RNA polymerase respectively, to express our target protein LL-37. To achieve this, we designed the DNA sequences of LL-37 to be inserted into the XbaI and XhoI sites of the pET-28a(+) vector (Figure 1.), transferred the recombinant plasmid into E.coli BL21(DE3) for protein expression.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 169
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]