Bacterial Cloning of Synthetic MTS1_03028

Both 5′ ends of the phosphorylated synthetic MTS1_03028 fragments optimized for E. coli codons were ligated with SmaI and pGEM7zf (+) and digested with T4 DNA ligase. The product was then transformed into Mach1-competent E. coli. A hundred microliters were spread onto LB agar containing 100 μg/ml ampicillin and statically incubated at 37°C overnight. The plasmids were then isolated and designated pGEM-03028.

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Ishida Y., Koretsune T., Ishiuchi E., Teshima M, & Ito M. (2023). A magnesium transporter is involved in the cesium ion resistance of the high-concentration cesium ion-resistant bacterium Microbacterium sp. TS-1. Frontiers in Microbiology, 14, 1136514.

Publication 2023

Agar Ampicillin Codons E coli Pgem Plasmids Smai T4 dna ligase

Corresponding Organization : Toyo University

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Variable analysis

independent variables

Ligation of 5' ends of phosphorylated synthetic MTS1_03028 fragments optimized for E. coli codons with SmaI and pGEM7zf (+)
Digestion with T4 DNA ligase

dependent variables

Transformation of the product into Mach1-competent E. coli
Growth of transformed E. coli on LB agar containing 100 μg/ml ampicillin
Isolation of plasmids designated pGEM-03028

control variables

Incubation temperature of 37°C
Overnight incubation period

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