Difference between revisions of "Part:BBa K4032102"

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Revision as of 09:58, 20 October 2021


lacI+RBS+α-gal+double terminator

Contents:

・The lac promoter (from BBa_R0010)

・The RBS (from BBa_B0034)

・The E.coli alpha-galactosidase gene melA ()

・The double terminator (from BBa_B0015)


Usage and Biology

This enzyme catalyzes the hydrolysis of terminal, non-reducing alpha-D-galactose residues in alpha-D-galactosides, including galactose oligosaccharides, galactomannans and galactolipids. https://www.uniprot.org/uniprot/P06720


Design

The lac promoter and the double terminator are added to No part name specified with partinfo tag., forming this part.

The lac promoter and the double terminator are used from BBa_J04450.


This part was created by In-Fusion method using BBa_J04450 and melA from E.coli K-12 as an insert.


1600px-T--Gunma--%CE%B1-galactosidase-stop-design-2.png


Fig.1 The plasmid design of K40320xx


Experiments

Time course BL21(DE3)

T--Gunma--%CE%B1-galactosidase-timecourse-BL21.png


Fig. 2 The growth of E.coli (BL21(DE3)) expressing of alpha-galactosidase

Pre-culture : 37 ℃, 16 h (130 rpm)

Culture : 37 ℃ (130rpm)

・Measuring OD600 every 4 hours


DH5α

T--Gunma--%CE%B1-galactosidase-timecourse-dh5%CE%B1.png


Fig. 3 The growth of E.coli (DH5α)) expressing of alpha-galactosidase

Pre-culture : 34 ℃, 16 h (130 rpm)

Culture : 37 ℃ (130rpm)

・Measuring OD600 every 4 hours


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2137
    Illegal AgeI site found at 2249
  • 1000
    COMPATIBLE WITH RFC[1000]