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Team:HKU-HKBU/Parts
From 2009.igem.org
Parts submitted to the Registry
This year, we constructed 50 and submitted to the Registry a total of 42 new standard biobricks with code numbers from K283000 to K283049. Please click [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=HKU-HKBU Biobricks from HKU-HKBU 2009] for their detailed information. Of these biobricks, FORTY-FOUR contain complete information of primary nucleaic acid sequence, description of function, authorship, safety notes, and acknowedgment of sources and references; TWO (BBa_K283003 and BBa_K283005) are the improved versions of the existing biobricks, BBa_K094102 and BBa_J36846, rescpectively; THIRTY-THREE, including SEVEN devices, (see Registry for more information) were demonstrated work as expected in our system.
After constructed a serial of intermediates, such as BBa_K283003, BBa_K283006, BBa_K283030, and so on, we eventually generated 4 USEFUL and FUNCTIONAL devices (BBa_K283000, BBa_K283001, BBa_K283002, and BBa_K283014), in terms of polar expression system and speed controlling system. In polar expression system, BBa_K283000 and BBa_K283001 were designed to display eGFP-streptavidin fusion protein on the cell surface. BBa_K283000 contains a signal peptide of lipoprotein (lpp) and five transmembrane domains of outer membrane protein A (ompA), while BBa_K283001 have the AIDA transmembrane domain. Figure 1 indicates the construction route of BBa_K283000. When expressed BBa_K283000 and BBa_K283001 in our Salmonalla strain YBS01 and E. coli strain YBE01, respectively, we clearly observed polar localization of eGFP-streptavidin protein (see our polar expression results).
To control the swimming speed of our engineered bacteria and simultaneously control the propelling force, we generated another two FUNCTIONAL devices BBa_K283002 and BBa_K283014. Figure 2 shows the assembly route of BBa_K283002. When it was expressed in strain YBE01, a proportional relation was observed between the CheZ expression level/bacterial swimming speed and the concentration of inducer (IPTG in this case) (see our speed control result).
