Difference between revisions of "Part:BBa K3034004"
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===Usage and Biology===
 
===Usage and Biology===
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To achieve lytic function, we usually express Lysep3-D8 protein. Combination of the inducible promoter with this part allows expression under specific conditions to inhibit the growth of ''E. coli''. We can also introduce it into ''E. coli'' BL21 (DE3) strain expression and use it as an "antibiotics" when a sufficient amount of protein is obtained.
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===Experiments===
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In order to verify the lytic effect of Lysep3-D8 protein, we cloned the corresponding coding gene into the vector induced by IPTG, and then induced the expression of this gene by adding IPTG with a final concentration of 0.5mM to obtain the intracellular lysis effect of Lysep3-D8 against ''E.coli'' DH5α(culture temperature 37 ̊C). At the same time, in order to verify the extracellular cleavage of ''E.coli'' BL21(DE3), we transformed the plasmid into ''E.coli'' BL21(DE3) and induced expression at 16 ̊C (Final concentration of IPTG was 0.5mM) for 16 hours to obtain a sufficient amount of protein. After that, we added the protein to E. coli DH5α cultured to logarithmic growth phase to verify the effect of extracellular lysis.
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===Results===
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We showed the growth of E. coli by measuring the growth curve of the Abs600 value of the bacterial liquid every hour. The results are as follows:
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Revision as of 08:41, 6 October 2019


The fusion protein is an enhanced lysin-> lysep3-D8

This lysin is formed by fusion of lysep3 and D8,which lyses bacteria from inside and outside the cell.The bactericidal spectrum of lysep3-D8 is broad, as it can lyse both gram-negative bacteria and gram-positive bacteria(14 E. coli strains, 3 P. aeruginosa strains, 1 Acinetobacter baumannii strain,and 1 Streptococcus strain).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 79
  • 1000
    COMPATIBLE WITH RFC[1000]