[DIYbio] Re: Exocytosis in Bacteria

Beta-Lactamase is a protein that is secreted by E coli.

ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA

The underlined peptide is cut off. Maybe it would work to just fuse your protein to it. Or leave 5-10 additional amino acids of the native N-terminus to make sure it is still recognized





On Thursday, February 5, 2015 at 11:40:18 PM UTC+1, Jacob Palumbo wrote:

Hey all,

I'm working on a project where it would be extremely useful to have bacteria (E. Coli specifically) exocytose proteins continuously, rather than grow a culture and lyse it the old fashion way. I've found a few papers on this so far (1, 2, 3), but they are all for Gram-positive bacteria, which E. Coli is not. If need be, I could switch over, but it would be nice to not have to. I do have some information the Sec pathway in Gram-negative bacteria, but it is somewhat general.

Anyone have an experience/insight into this?

Thanks,
Jake