Team:HKU-HKBU/Parts

From 2009.igem.org

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=Parts submitted to the Registry=
=Parts submitted to the Registry=
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'''This year, we constructed and submitted to the Registry a total of 42 new standard biobricks with code number 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, XX () contain complete information of primary nucleaic acid sequence, description of function, authorship, safety notes, 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; XX (BBa_) were demonstrated work as expected in our system.'''
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'''This year, we constructed 50 and submitted to the Registry of Standard Parts 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 on primary nucleic 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.'''
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BBa_K283000 and BBa_K283002 are genetic circuits for polar expression and speed control respectively.
 
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After constructing a serial of intermediates, such as BBa_K283003, BBa_K283006, BBa_K283030, and so on, we eventually generated ''four'' '''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 ([https://2009.igem.org/Team:HKU-HKBU/Polar_Expression_Results#Polar_Expression see our polar expression results]).   
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===Assembly route of BBa_K283000:===
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[[Image:K283000new.jpg|center|thumb|700px|'''Figure 1.''' Assembly route of BBa_K283000]]
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To simultaneously control the swimming speed and propelling force of our engineered bacteria , 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) ([https://2009.igem.org/Team:HKU-HKBU/Speed_Control_Results#Regulation_of_cheZ_expression see our speed control result]).
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[[Image:K283000new.jpg|center|caption]]
 
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[[Image:Assembly of BBa K283002new.jpg|center|thumb|700px|'''Figure 2.''' Assembly route of BBa_K283002]]
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===Assembly route of BBa_K283002:===
 
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[[Image:Assembly of BBa K283002new.jpg|center|caption]]
 
{{Team:HKU-HKBU/footer}}
{{Team:HKU-HKBU/footer}}

Latest revision as of 20:27, 21 October 2009

Parts submitted to the Registry

This year, we constructed 50 and submitted to the Registry of Standard Parts a total of 42 new standard biobricks with code numbers from K283000 to K283049. Please click Biobricks from HKU-HKBU 2009 for their detailed information. Of these biobricks, FORTY-FOUR contain complete information on primary nucleic 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 constructing a serial of intermediates, such as BBa_K283003, BBa_K283006, BBa_K283030, and so on, we eventually generated four 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).


Figure 1. Assembly route of BBa_K283000



To simultaneously control the swimming speed and propelling force of our engineered bacteria , 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).


Figure 2. Assembly route of BBa_K283002



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