Drosophila P-Element Transposons
Optogenetics requires cell specific gene expression to manipulate neural activity in a specific group of neurons. In Drosophila, transgenic methods are widely used for this purpose.
The P-element is a Drosophila specific transposon for mutagenesis because it has a reverse repeat of 31 base pairs at both ends. It also contains a DNA sequence encoding transposase, which recognizes 31 pairs of sequences and can cut and insert DNA into both ends of the genome.
Creative BioMart is now able to provide custom Drosophila strain construction service. The service includes the injection, generation and identification of transformants.
Technology Principles
A vector containing foreign gene was prepared and the sequence was sandwiched between 31 base pair reverse repeats. Within one hour after the egg was laid, the vector containing transposase was injected into the fertilized egg of the strain lacking endogenous P-element transposon. In embryos, exogenous DNA is randomly inserted into the genomes of some cells under the action of transposase. DNA injected into the embryo spreads throughout the embryo and can be integrated into any nucleus, including the nucleus of hypothetical germ cells. Therefore, transgenic animals with stable foreign genes in all cells can be obtained in the offspring of germ cells undergoing integration.
Overview of the Workflow
According to our equipment platform, we will generate transgenic fruit flies for optogenetic research according to the following process.
|
Service Steps |
Description |
| Drosophila strain preparation |
Unless otherwise required, we will select the white mutant w1118 containing the Mini White reporter gene as the original Drosophila strain. Amplification of the white mutant began a few weeks before the planned injection. |
| Embryo injection |
We will provide five independent transformation systems unless the insertion amount is large (i.e. greater than 13 KB). |
| Establishment and mapping of transgenic lines |
DNA was injected into w1118 embryos, the surviving larvae were cultured to adulthood and hybridized with G0 Drosophila (> 100 individuals).
After the identification and collection of transformants, the collected transformed fruit flies (at least 3 independent strains) were sent to the customer, and the original vials were kept in the facility for two weeks as standby. |
Available Schemes at Creative BioMart
| Scheme 1 |
Inject 100 embryos |
Provide surviving larvae |
| Scheme 2 |
Inject 100 embryos and provide unbalanced G2 transformants. |
> 4 weeks |
| Scheme 3 |
Inject 100 embryos and provide balanced lines. |
> 88 weeks |
*Scheme 1 is injection only. we will inject into white1118 embryos unless you request otherwise.
Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.
