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From 2014.igem.org

As a student team, we highly appreciated recommendations and feedback from experts of different scientific areas. Especially for a scientific project like the iGEM competition, it was valuable to unclose new perspecives and assess feasibility of our project ideas.

Experts on Inteins

= Prof. Dr. Henning D. Mootz (WWU Münster)

When we started our literature research on the use of inteins for posttranslational processing we realized the great contributions of Prof. Dr. Henning D. Mootz to this field of research. Dr. Henning Mootz holds a Professorship in Biochemistry and Biotechnology at the Institute of Biochemistry WWU Münster (Germany). In our conversation he gave a nice overview about the most prominent inteins that are used in biotechnology or research and highlighted their special features. We further discussed about the use of inteins for our aim of circularization and appreciated his offer to send us appropriate intein constructs very much. Prof. Dr. Henning Mootz highlighted that until now the induction of inteins cannot be regulated by an in vivo expressible biological mechanism. Therefore, he strongly recommended to pursue this very interesting approach. Moreover, we talked with him about the oligomerization of multidomain proteins to allow expression of their components in E.coli. When talking about the circularization of large proteins, Prof. Dr. Mootz emphasized that efficiency of this approach cannot be predicted due to the complex nature of protein folding. Nevertheless, he commented that proteins have to be imagined as dynamic entities that can potentially be circularized. Nevertheless, efficiency and preservation of the protein function would have to be part of our experiment.

At this point we would again like to thank Prof. Dr. Henning D. Mootz for providing his constructs and sharing his great expertise with us.

PD Dr. Matthias Mayer (University Heidelberg)

The iGEM team Heidelberg met Dr Matthias Mayer, group leader in the center for molecular biology in Heidelberg (ZMBH), for expert counsel. Dr Meyer already knew about intein splicing and various applications and advantages that were proved effective in literature. Together we philosophized about new ideas of intein utilization and our DNMT1 project. He thinks that circularizing DNMT1 would be a risky project, since the increase of thermo stability cannot be predicted. Anyhow if the stability of DNMT1 increases, it would be a break- through not only in terms of a new intein application, but also regarding the opportunities that arise with a heat-stable DNMT1. Matthias Mayer was also fascinated about the idea of a linker software, that calculates the best linker to connect the ends of a protein and evaluates its rigidity to further increase thermo stability. If we manage to set up this software, he would definitely use it!