Experiment Parts

Abstract

In terms of genetic engineering, the main goal of the project is to use E. coli BL21 to express two interacting proteins from Homo sapiens, MST2 and STRN3. The interaction of these two proteins leads to dysregulation of the Hippo pathway, which in turn induces gastric, intestinal, liver, and breast cancer, among others. We cloned the MST2 and STRN3 genes from the human genome and co-expressed them by fusing GST- and HST-tagged proteins, respectively, to facilitate subsequent purification. After the software predicted the three amino acid sites where MST2 interacts with CX6258, we mutated them to obtain MST2 (mutant) for pull-down experiments to verify the accuracy of the predicted sites.

To achieve the above goal, we successfully constructed 7 parts. BBa_K4889000, BBa_K4889001, and BBa_K4889003 are basic parts for MST2 1-308, STRN3 64-145, and MST2 1-308 (mutant) genes, respectively. BBa_K4889002 is a GST-tagged protein gene. BBa_K4889004, BBa_K4889006, and BBa_K4889007 are devices for co-expressing MST2 1-308, MST2 1-308 (mutant) and STRN3 64-145 genes with lac promoter and corresponding tag.