Sperm-mediated transformation of the honey bee, Apis mellifera.
Our primary objective was to identify techniques to transform the genome of the honey bee (Apis mellifera) with foreign DNA constructs. The strategy we adopted was to linearize foreign DNA and introduce it with sperm during the instrumental insemination of virgin queen honey bees. We analysed extracts from larvae within the same cohort and isolated the predicted fragment by means of PCR amplification of genomic DNA. Larvae that carried the construct also expressed the introduced DNA. We propagated several transgenic lines for up to three generations, which demonstrates its heritability. Once carried by a queen, the construct can be detected in that queen's larvae over several months. However, there was no evidence of integration of the construct, at least as determined by genomic Southern analysis. Nevertheless, this demonstrates the general viability of the technique for introduction of DNA, and it should be augmented by further use of transposable elements that enhance integration.
Robinson
K O
KO
Molecular, Cellular, and Developmental Biology Program, The Ohio State University, Columbus, OH 43210, USA.
Ferguson
H J
HJ
Cobey
S
S
Vaessin
H
H
Smith
B H
BH
D000818:Animals; D030801:Animals, Genetically Modified; D001516:Bees / Q000235:genetics*; D015139:Blotting, Southern; D005260:Female; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D010957:Plasmids; D016133:Polymerase Chain Reaction; D013094:Spermatozoa / Q000502:physiology*; D014170:Transformation, Genetic
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins