Difference between revisions of "Part:BBa K4032102"

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Contents:
 
Contents:
  
・The lac promoter (from <partinfo>BBa_R0010</partinfo>)
+
・The lac promoter from <partinfo>BBa_R0010</partinfo>
  
・The RBS (from <partinfo>BBa_B0034</partinfo>)
+
・The RBS from <partinfo>BBa_B0034</partinfo>
  
・The α-galactosidase gene ''melA'' from ''E. coli'' <partinfo>BBa_K4032000</partinfo>
+
・The α-galactosidase gene ''melA'' from ''E. coli'' <partinfo>BBa_K4032000</partinfo>
  
・The double terminator (from <partinfo>BBa_B0015</partinfo>)
+
・The double terminator from <partinfo>BBa_B0015</partinfo>
  
  
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===Usage and Biology===
 
===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 (see info [https://www.uniprot.org/uniprot/P06720 from UniProt EC:3.2.1.22]).
+
This enzyme catalyzes the hydrolysis of terminal, non-reducing alpha-D-galactose residues in alpha-D-galactosides, including galactose oligosaccharides, galactomannans and galactolipids. For more information, [https://www.uniprot.org/uniprot/P06720 UniProt EC:3.2.1.22].
  
  

Revision as of 13:37, 21 October 2021


lacI+RBS+α-gal+double terminator

Contents:

・The lac promoter from BBa_R0010

・The RBS from BBa_B0034

・The α-galactosidase gene melA from E. coli BBa_K4032000

・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. For more information, UniProt EC:3.2.1.22.



Design

The lac promoter and the double terminator are added to BBa_K4032005, forming this part.

The lac promoter and the double terminator are from BBa_J04450.


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


800px-T--Gunma--%CE%B1-galactosidase-stop-design-2.png

Fig.1 The plasmid design of K40320102


Experiments

Time course BL21(DE3)

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


Fig. 2 The growth of E. coli (BL21(DE3)) expressing α-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 α-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]