Difference between revisions of "Part:BBa K1949001:Design"
(→Design Notes) |
|||
| Line 8: | Line 8: | ||
===Design Notes=== | ===Design Notes=== | ||
| − | We applied a point mutation to the P<i>cold</i> in order to be easy to clone it. We changed the | + | We applied a point mutation to the P<i>cold</i> in order to be easy to clone it. We changed the 177th base C to A. An article which we referred describes as stated below.<br> |
“We found occasionally that C–A base substitution at nucleotide number 532 in the 5’-UTR region (nucleotide number is according to GenBank Accession No. M30139) diminished this growth inhibition effect (data not shown), allowing easy cloning of the genes of interest into MCS. We do not know why this mutation diminished growth inhibition effect but the mutation did not affect productivity of proteins (data not shown).”<br> | “We found occasionally that C–A base substitution at nucleotide number 532 in the 5’-UTR region (nucleotide number is according to GenBank Accession No. M30139) diminished this growth inhibition effect (data not shown), allowing easy cloning of the genes of interest into MCS. We do not know why this mutation diminished growth inhibition effect but the mutation did not affect productivity of proteins (data not shown).”<br> | ||
Revision as of 16:56, 10 October 2016
Pcold-gfp
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 308
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 957
Design Notes
We applied a point mutation to the Pcold in order to be easy to clone it. We changed the 177th base C to A. An article which we referred describes as stated below.
“We found occasionally that C–A base substitution at nucleotide number 532 in the 5’-UTR region (nucleotide number is according to GenBank Accession No. M30139) diminished this growth inhibition effect (data not shown), allowing easy cloning of the genes of interest into MCS. We do not know why this mutation diminished growth inhibition effect but the mutation did not affect productivity of proteins (data not shown).”
Nakashima,N and Tamura,T. 2004. Cell-free protein synthesis using cell extract of Pseudomonas fluorescens and CspA promoter. Biochemical and Biophysical Research Communications 319 (2004) 672