Template:SBB09 20154

Sadao Ota

Ag43 Display
Source: E. coli MG1655 genomic DNA You are going to make an antigen 43 autotransporter display system based on this reference: PMID: 12107137. The gene encoding Ag43 is called flu, and the DNA sequence is here. You should also look at the protein sequence at YP_026164 which includes some annotation of the domains within the protein. The actual transporter portion of the protein is: Region         738..1017 /region_name="Autotransporter" Upstream of this sequence is one of the passenger domains: Region         517..707 /region_name="PL2_Passenger_AT"

For your first part, you should make the part corresponding to amino acids 708-1017. For your second part, you should make the part corresponding to amino acids 517-1017.

Due to the long length of the second part, you will not sequence your plasmid completely. Just sequence the ends with ca998 and G00101 to confirm the first and last 500bp.