Module 2, Lab 06 - Ligation and transformation (GAPC gene from Arabidopsis and pJet1.2 plasmid)

Today we purified the PCR products from the nested PCR lab (GAPC gene from Arabidopsis) and used them to genetically transform E. coli.

The lab was divided in three main steps

• Ligation (of GAPC gene on to the pJet1.2 plasmid)
• Preparation of competent cells
• Genetic transformation of E. coli

We spent most of the lab manipulating bacteria to make them competent (i.e. get them ready to uptake extracellular DNA). Once this was achieved, the GAPC gene from Arabidopsis, obtained via nested PCR, was ligated to the pJet1.2 plasmid.
The plasmid was then used to genetically transform E. coli, which were spread on LB agar/Amp/IPTG plates and incubated.

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Lecture, chapter 09 - Gene regulation in prokaryotes

Today we lectured for a little over one hour, as a replacement for the lab we could not do, because of not having transformed bacteria to "play" with.

We finished chapter 9 on gene regulation in prokaryotes. We discussed the concepts of positive and negative regulation, including the role of (specific) activators and repressors, and global regulators. We studied how the Lac operon works in E. coli, and set the stage for starting with gene regulation in eukaryotes.

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Lecture, chapter 09 - Gene regulation in prokaryotes

We started the fourth stop in our "road map" (how genes are regulated).

We introduced chapter 09, on gene regulation, by highlighting the importance of the process for both eukaryotic and prokaryotic cells. We also mentioned the different steps during the process of information transfer at which regulation can take place, transcription being the most common one.

Note: Because the results of the transformation lab were negative (E. coli genetic transformation failed) we will lecture tomorrow.

• Section 01: 8:45 am
• Section 02: 10:00 am
• (Any one who wants to attend the opposite section is welcome)

Information on how to obtain the results from the transformation lab will be posted on an update in that exercise's blog entry.

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Lecture, chapter 7 - Protein structure and function

We finished chapter 7...

We discussed the features that allow certain proteins to bind to DNA and also what of their most common structural motifs are. We closed by briefly describing what protein denaturation is, how it differs from degradation, and what are the most common denaturing agents.

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Lecture, chapter 7 - Protein structure and function

Today we covered most of chapter 7, on protein structure and function.

We discussed the properties a protein has given the characteristics amino acids provide to the whole structure and how they actually affect protein function. We also listed different protein categories according to different functions they may have.

Tomorrow we will talk about how proteins can recognize DNA sequences to bind to them.

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