Kanamycin in Cereal Biotechnology: A Screening or a Selectable Marker?

Malik Shuja,Muhsin Jamal,Muhammad Imran,Hassan Riaz

doi:10.32350/bsr.0301.i

Abstract

Kanamycin is a widely used selection agent in dicot-plant genetic transformation systems. In monocots, however, it does not seem to be effective as it has no or minimal effect on the normal growth of non-transformed plants. Kanamycin was previously demonstrated to bleach the pigments of the non-transgenic plants. This may yield the idea that kanamycin can be used as an effective screening marker rather than a selectable marker in monocots. Copyright (c) 2021 Malik Nawaz Shuja, Hasan Riaz, Muhsin Jamal, Muhammad Imran

Highlights

A kanamycin scavenging gene nptII, expressed by Escherichia coli transposon Tn5, encodes neomycin phosphotransferase II enzyme – known as aminoglycoside 3'phosphotransferase II [5, 6, 7]. This enzyme transfers the γ-phosphate group of ATP to the 3'-hydroxyl group of the amino-hexose residue of the aminoglycoside antibiotics that results in detoxification
In a genetic transformation setting, kanamycin is supplemented to the growth medium in a concentration that may inhibit untransformed cells from regeneration
Kanamycin does not seem to be mobile in the vascular tissue; rather, it seemingly diffuse through the plant tissue via intercellular spaces

Summary

Introduction

A kanamycin scavenging gene nptII (or neo), expressed by Escherichia coli transposon Tn5, encodes neomycin phosphotransferase II enzyme – known as aminoglycoside 3'phosphotransferase II [5, 6, 7]. This enzyme transfers the γ-phosphate group of ATP to the 3'-hydroxyl group of the amino-hexose residue of the aminoglycoside antibiotics that results in detoxification. This ATP-dependent phosphorylation allows protein synthesis, thereby resisting the specific binding of antibiotics to ribosomes. The kanamycin A, B, and C, neomycin, paromomycin, and geneticin – aminoglycoside antibiotics containing 3'OH group – are substrates of NPTII [8].

Results

Conclusion