01Abr - 2024
Homologous recombination cloning: Tips, tricks and advantages
05:00 PM - 07:00 PM|Dr. Rafael Martinez Gallegos|Developmental Biology, Centre for Organismal Studies (COS), Heidelberg University, Germany|Invitado por: Joseph Dubrovsky
Seminario
Using the endogenous homologous recombination of living organisms for cloning (HR-cloning) was first described in yeast and eventually in bacteria. Although not so widespread, this method allows the straightforward engineering of fusion proteins with the ability of exchanging elements from 1 nt to several kb within a plasmid. Therefore it can be used to manipulate codons or transcription factor target sequences as well as to study regulatory regions like full promoters or UTRs. Likewise, reporter markers can be exchanged having either N- or C-terminal variants. It also offers the possibility of engineering customized plasmids providing a different strategy to overcome difficulties in the cloning and editing of challenging DNA fragments. The manipulation steps are minimized and any recA1 bacteria strain is suitable for it. This method has been tested with common and customized plasmids with a range up to 15 kb. Its simplicity allows the implementation in any molecular biology laboratory without the requirement of any commercial Kit. I will discuss examples of HR-cloning on a wide variety of organisms, from plants to Drosophila.
Actualizado 2024-03-11 17:26:35
03-Junio-2024 al 03-Junio-2024
12:00 PM
Dr. Takashi Yamamoto
12:00 PM
Dr. Takashi Yamamoto
I am planning to cover some of the topics that I was focused during my career in the field of Bacillus thuringiensis: 1. Discovery of new Bt insecticidal proteins. 2. Structure and mode of action of Cry2A. 3. Mode of action of other Cry Proteins. 3. Engineering Cry proteins for improved insect control, by different technologies such as Domain Swapping, Supporting Technologies, DNA Shuffling, Saturation Mutagenesis. 4. Bacillus subtilis Spore Display.