Difference between revisions of "Part:BBa K533012:Experience"

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<I>Tsinghua</I>
 
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We transform Top10 competent cells with 1ng of such plasmid and incubated the petri plates at 37 centigrade. At most one colony was formed on each plate, indicating that ccdB is working and killing the non-resistant cells.
 
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Revision as of 18:56, 28 October 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K533012

This is a backbone derived from pSB1C3 and confers the host chloramphenicol resistance.

E/X/S/P MCS is present in this backbone and thus can be used for convenient parts assembly.

What is different for this backbone is that it expresses a ccdB gene, which will cause the death of commonly used E. coli strains.

Hence, do NOT use common competent cells when this plasmid is involved. Instead, use a ccdB resistance strain.

A mutated I-SceI site is also present in the plasmid, which allows for elimination of heavily mutated vectors in I-SceI expressing strains.

This year, we tried to construct an E. coli strain expressing ccdB resistance gene and I-SceI endonuclease from its chromosome and wanted to use this new strain as the engineering strain for all the biobricks. Due to the limited time, we did not construct this strain. Any teams interested in enhancing safety level might consider this undertaking.

User Reviews

UNIQ2413dc3f5a756bbf-partinfo-00000000-QINU

BBa_K533012 5 Not understood Tsinghua

We transform Top10 competent cells with 1ng of such plasmid and incubated the petri plates at 37 centigrade. At most one colony was formed on each plate, indicating that ccdB is working and killing the non-resistant cells.

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UNIQ2413dc3f5a756bbf-partinfo-00000002-QINU