Difference between revisions of "Part:BBa K4032102"
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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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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]