Difference between revisions of "Part:BBa C0261:Experience"

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We have sequenced the part and compared the result with the data in NCBI. Although many nucleic acid of DNA sequence are different, the amino acid sequence is the same. We use PCR to get the LuxI gene and build more complicated parts.
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We modify this part this year. Firstly, we delete the LVA tag on the 3' end. This can improve the production of AHL but may have some bad influence on the E.coli's metabolism. Secondly, we add the His-tag on the N-terminal of protein. With the help of it, we can easily purify the protein using Ni-column. Also, you can use anti-histag antibody to detect the protein's expression in western blot. Finally, we add some enzyme cutting site which can facilate the construction.
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Our new part is Bba_K766000.

Revision as of 12:08, 26 September 2012

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_C0261

I am the team member of Tsinghua. In theory, LuxI is a C6HSL synthase so we use this part to produce AHL. This molecule can move across the membrane freely so that it can be used as a signal to transmit information.

In practice, we find some tricks about how to use this part, which will be talked in experience below.

User Reviews

UNIQaca3446a6e8d969b-partinfo-00000000-QINU

Antiquity

This review comes from the old result system and indicates that this part did not work in some test.

UNIQaca3446a6e8d969b-partinfo-00000002-QINU We have sequenced the part and compared the result with the data in NCBI. Although many nucleic acid of DNA sequence are different, the amino acid sequence is the same. We use PCR to get the LuxI gene and build more complicated parts.

We modify this part this year. Firstly, we delete the LVA tag on the 3' end. This can improve the production of AHL but may have some bad influence on the E.coli's metabolism. Secondly, we add the His-tag on the N-terminal of protein. With the help of it, we can easily purify the protein using Ni-column. Also, you can use anti-histag antibody to detect the protein's expression in western blot. Finally, we add some enzyme cutting site which can facilate the construction.

Our new part is Bba_K766000.