Difference between revisions of "Part:BBa K4032102"
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Contents: | Contents: | ||
| − | ・The lac promoter | + | ・The lac promoter from <partinfo>BBa_R0010</partinfo> |
| − | ・The RBS | + | ・The RBS from <partinfo>BBa_B0034</partinfo> |
| − | ・The | + | ・The α-galactosidase gene ''melA'' from ''E. coli'' (<partinfo>BBa_K4032000</partinfo>) |
| − | ・The double terminator | + | ・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. https://www.uniprot.org/uniprot/P06720 | + | 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]. |
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===Design=== | ===Design=== | ||
| − | The lac promoter and the double terminator are added to <partinfo> | + | The lac promoter and the double terminator are added to <partinfo>BBa_K4032005</partinfo>, forming this part. |
| − | The lac promoter and the double terminator are | + | The lac promoter and the double terminator are from <partinfo>BBa_J04450</partinfo>. |
| − | This part was created by In-Fusion method using <partinfo>BBa_J04450</partinfo> and melA from E.coli | + | This part was created by In-Fusion method using <partinfo>BBa_J04450</partinfo> and melA from ''E. coli'' as an insert. |
https://2021.igem.org/wiki/images/thumb/1/1f/T--Gunma--%CE%B1-galactosidase-stop-design-2.png/800px-T--Gunma--%CE%B1-galactosidase-stop-design-2.png | https://2021.igem.org/wiki/images/thumb/1/1f/T--Gunma--%CE%B1-galactosidase-stop-design-2.png/800px-T--Gunma--%CE%B1-galactosidase-stop-design-2.png | ||
| − | Fig.1 The plasmid design of | + | Fig.1 The plasmid design of BBa_K4032102 |
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| − | Fig. 2 The growth of E.coli (BL21(DE3)) expressing | + | Fig. 2 The growth of ''E. coli'' (BL21(DE3)) expressing α-galactosidase |
| − | Pre-culture : 37 ℃, 16 h (130 rpm) | + | Pre-culture: 37 ℃,16 h (130 rpm) |
| − | Culture : 37 ℃ ( | + | Culture: 37 ℃ (130 rpm) |
･Measuring OD600 every 4 hours | ･Measuring OD600 every 4 hours | ||
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| − | Fig. 3 The growth of E.coli (DH5α)) expressing | + | Fig. 3 The growth of ''E. coli'' (DH5α)) expressing α-galactosidase |
| − | Pre-culture : 34 ℃, 16 h (130 rpm) | + | Pre-culture: 34 ℃,16 h (130 rpm) |
| − | Culture : 37 ℃ ( | + | Culture: 37 ℃ (130 rpm) |
･Measuring OD600 every 4 hours | ･Measuring OD600 every 4 hours | ||
Latest revision as of 02:45, 22 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.
Fig.1 The plasmid design of BBa_K4032102
Experiments
Time course BL21(DE3)
Fig. 2 The growth of E. coli (BL21(DE3)) expressing α-galactosidase
Pre-culture: 37 ℃,16 h (130 rpm)
Culture: 37 ℃ (130 rpm)
･Measuring OD600 every 4 hours
DH5α
Fig. 3 The growth of E. coli (DH5α)) expressing α-galactosidase
Pre-culture: 34 ℃,16 h (130 rpm)
Culture: 37 ℃ (130 rpm)
･Measuring OD600 every 4 hours
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2137
Illegal AgeI site found at 2249 - 1000COMPATIBLE WITH RFC[1000]