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NPTII Assays for Measuring Gene Expression and Enzyme Activity in Transgenic Plants

 Last Update: 2020-11-17
 Source: Internet
 Author: User

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black marker pen

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The stable introduction of genes into plants through genetic engineering normally necessitates the use of a selectable marker, especially when the transformation frequency is low (e.g., 1.0 � 10
−3
to 10
−6
). Marker genes enable quantification of both the transformation efficiency and gene expression (
1
). The most commonly used selectable marker is the gene from Transposon 5 (Tn5) of
Escherichia coli
K12
(aphA2)
(
2
), which encodes aminoglycoside 3-phosphotransferase II (APH [3] II, Chemical Abstracts Registry number 58943-39-8) activity. This enzyme, also known as neomycin phosphotransferase II (NPTII), inactivates by phosphorylation the sugar-containing antibiotics, neomycin, kanamycin, geneticin (G418), and paromomycin. To date, the gene has been introduced into over 30 plant species. (For a detailed description of the
nptII
gene,
see
Chapter 1. )

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