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Foreign gene induction into Bombyx mori egg by electroporation
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/290764
Electroporation is well known today as a powerful transfection technique and is useful for the study of gene expression. The advantage of the electroporation method is that large quantity of silkworm (Bombyx mori) eggs can be transformed in a very short time. However, how to use it for introducing foreign gene into silkworm eggs needs systematical investigation. In our silkworm transgenesis program, we needed an efficient technique to evaluate the functionality of transgenes before their injection into eggs. The goal of this experiment was to find an alternative efficient method of generating transgenic silkworm eggs using a commercially available electroporation device. The Gene Pulser Xcell (Bio-Rad Laboratories, USA) were used. In contrast to other electroporation devices, which are based on a single pulse with exponential decay or square wave technology. We investigated pigmentation-rate and hatching-rate of the silkworm eggs of electroporation. We used foreign gene LacZ, EGFP, Ds-red induced vector system with selection marker for transgenic silkworm. The LacZ gene in 3rd instar larva DNA can be detected by β-galactosidase stain. During these technical studies we found that optimizing parameters such as electrical voltage, number of pulses and their frequency, and conductivity of the buffer was important. These results confirmed that electroporation is available technique for transfecting B. mori egg.
