Difference between revisions of "Part:BBa K4032102"

Revision as of 08:46, 20 October 2021

lacI+RBS+α-gal+double terminator

Contents:

・The lac promoter https://parts.igem.org/Part:BBa_R0010

・The RBS https://parts.igem.org/Part:BBa_B0034

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

・The double terminator https://parts.igem.org/Part: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 BBa_K40320xx, forming this part.

The lac promoter and the double terminator are used from BBa_J04450 (https://parts.igem.org/Part:BBa_J04450) .

This part was created by In-Fusion method using BBa_J04450 (https://parts.igem.org/Part:BBa_J04450) and melA from E.coli K-12 as an insert.

Fig.1 The plasmid design of K40320xx

Experiments

Time course BL21(DE3)

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

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