Team:TUDelft/8 July 2009

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='''8th July'''=
='''8th July'''=
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=='''Conjugation'''==
=='''Conjugation'''==
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Today, we made an unbelievable discovery: we found out that the conjugation between two cells with the same plasmids has been done before by the [http://parts.mit.edu/igem07/index.php/Peking_Hop-Count Peking team from iGEM 2007]. In conjugation, the Tra family proteins are present on the F plasmid and are responsible for establishing the connection between the conjugating cells. The Peking team took traI out of the F plasmid, and implemented it into a miniF plasmid to allow only cells who contain the miniF plasmid would be able to conjugate with other cells. The problem that Peking had with using traI was that it had several restriction sites that were not allowed by the iGEM standards. Nevertheless, we looked today at a small part from the traI that had crucial genes for conjugation and did not contain illegal segments. We are still not sure whether this gene will be removed properly from the F plasmid and function well within the miniF, so we are still trying to consider some alternative tra genes in the F plasmid that are important to conjugation.
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Today, we worked on the entries to the Parts Registry, where we used some parts from the Peking '07 iGEM team and some parts from NCBI. After writing down the sequences for trbC (pilin) of IncP beta R751 plasmid, trbK (entry exclusion) of IncP beta R751 plasmid, I-PpoI homing endonuclease site, I-SceI homing endonuclease site and I-PpoI homing endonuclease into the registry, we used the software [http://www.vectorcore.pitt.edu/upgene/upgene.html Upgene] to optimize for the codons, and the optimized sequences were inserted into the Parts Registry.
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Latest revision as of 14:27, 25 July 2009

Lab Notebook

July
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August
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8th July


Conjugation

Today, we worked on the entries to the Parts Registry, where we used some parts from the Peking '07 iGEM team and some parts from NCBI. After writing down the sequences for trbC (pilin) of IncP beta R751 plasmid, trbK (entry exclusion) of IncP beta R751 plasmid, I-PpoI homing endonuclease site, I-SceI homing endonuclease site and I-PpoI homing endonuclease into the registry, we used the software [http://www.vectorcore.pitt.edu/upgene/upgene.html Upgene] to optimize for the codons, and the optimized sequences were inserted into the Parts Registry.


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