Difference between revisions of "Part:BBa K4015001"

Revision as of 10:15, 20 October 2021

Lcp1VH2

The Lcp1VH2 is a strain of latex clearing protein extracted from Gordonia polyisoprenivorans VH2, a species of Gordoniaceae, which is of interest because of its ability to damage natural rubber. Latex clearing protein was first described by Rose (Rose et al., 2005) and the synthesis of Lcp1VH2 into e.coli was first described by Hiessl et. al (Hiessl et al., 2014). The Lcp1VH2 belongs to oxygenate, which can catalyse the extracellular cleavage of poly (cis-1,4-isoprene) through adding two oxygen molecules to the chemical bond where the polyisoprene is attached(Ilcu et al., 2017). The molecular mass of latex clearing protein is 42 kDa.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 508
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 127
Illegal NgoMIV site found at 485
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 112
Illegal SapI.rc site found at 100

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 ===Usage and Biology===
+In previous studies, we have discovered that lcp1VH2 was usually inserted into the pET23a plasmid and subsequently transferred into the C41 strain for expression. We tried that approach, but the bands of lcp1VH2 were indistinguishable and could not tell whether it was expressed in E. coli C41.
+In order to obtain a viable expression system, we tested four expression protocols using existing strains on hand (E.coli C41 and E.coli BL21) with vectors (pet23a and pet28a): E.coli C41 pET23a:: Lcp1VH2, E.coli C41 pET28a:: Lcp1VH2, E. coli BL21 pET23a:: Lcp1VH2, E. coli BL21 pET28a:: Lcp1VH2 and E. coli BL21 pET28a:: cp1VH. coli BL21 pET23a:: Lcp1VH2 and E. coli BL21 pET28a:: Lcp1VH2. as shown in Fig1, we can see that E. coli BL21 pET23a:: Lcp1VH2 is the best system for Lcp1VH2 expression: compared to the control, we can see a clear induction band and the Lcp1VH2 expression and water solubility were excellent.
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