S. Gendreau et al., TRANSIENT EXPRESSION OF A LUCIFERASE REPORTER GENE AFTER BALLISTIC INTRODUCTION INTO ARTEMIA-FRANCISCANA (CRUSTACEA) EMBRYOS, Aquaculture, 133(3-4), 1995, pp. 199-205
Using the PDS 1000/He ballistic gene transfer apparatus from BioRad, A
rtemia franciscana embryos were bombarded with high velocity gold micr
oparticles coated with a circular plasmid containing a luciferase repo
rter gene placed under the control of the Drosophila melanogaster hsp
70 promoter (3000 to 10000 embryos per shot). Even for the most drasti
c ballistic parameters investigated on dechorionated cysts (rupture di
sk, 1800 psi; distance D between the stopping screen and the embryos,
35 mm), no significant luciferase activity was detected after heat sho
ck (30 min, 41 degrees C). After bombarding protruding embryos obtaine
d 20 h after dechorionated cyst rehydration, the best result in terms
of embryo survival and luciferase activity was obtained for D = 35 mm
with a 450-psi rupture disk. For these parameters, significant lucifer
ase activity was observed 12 and 24 h after bombardment, but not at 48
h. In this particular model, the data suggest transient luciferase ex
pression, with a peak of luciferase activity within the 12 h following
bombardment. These results indicate that the ballistic technique is u
seful for studying transgene promoter efficiency in crustaceans and su
ggest its utility for attempting the production of transgenic animals.