Difference between revisions of "Part:BBa K4119008"

Revision as of 15:44, 11 October 2022

Plac-ΔperR

The upper and lower homology arms (HR-UP, HR-DOWN) were constructed at about 500 bp upstream and downstream of the PerR sequence of the Clostridium tyrobutyricum genome, plus a lactose-inducing promoter and a spacer sequence derived from the PerR , generating the CRISPR array .Then inserted it into the pMTL82151 plasmid, the successful plasmid is transformed into E. coli CA434, and introduced into Clostridium tyrobutyricum, by lactose induction to make the spacer hit the perR sequence on the target genome, thereby utilizing the endogenous CRISPR-Cas system knocks the perR out , eliminates the impediment effect of perR on the growth of Clostridium butyrate, thereby achieving aerobic growth of Clostridium tyrobutyricum.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 368
Illegal PstI site found at 1597
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 368
Illegal PstI site found at 1597
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 368
Illegal BglII site found at 428
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 368
Illegal PstI site found at 1597
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 368
Illegal PstI site found at 1597
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4119008 short</partinfo>
-    The upper and lower homology arms (HR-UP, HR-DOWN) were constructed at about 500 bp upstream and downstream of the PerR sequence of the Clostridium tyrobutyricum genome, plus a lactose-inducing promoter and a spacer sequence derived from the PerR , generating the CRISPR array .Then inserted it into the pMTL82151 plasmid, the successful plasmid is transformed into E. coli CA434, and introduced into Clostridium tyrobutyricum, by lactose induction to make the spacer hit the perR sequence on the target genome, thereby utilizing the endogenous CRISPR-Cas system knocks the perR out , eliminates the impediment effect of perR on the growth of Clostridium butyrate, thereby achieving aerobic growth of Clostridium tyrobutyricum.
+The upper and lower homology arms (HR-UP, HR-DOWN) were constructed at about 500 bp upstream and downstream of the PerR sequence of the Clostridium tyrobutyricum genome, plus a lactose-inducing promoter and a spacer sequence derived from the PerR , generating the CRISPR array .Then inserted it into the pMTL82151 plasmid, the successful plasmid is transformed into E. coli CA434, and introduced into Clostridium tyrobutyricum, by lactose induction to make the spacer hit the perR sequence on the target genome, thereby utilizing the endogenous CRISPR-Cas system knocks the perR out , eliminates the impediment effect of perR on the growth of Clostridium butyrate, thereby achieving aerobic growth of Clostridium tyrobutyricum.
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