Team:SUSTech Shenzhen/Test3D
Description
View
Contents
Overview
Why Caenorhabditis elegans?
What is MiniMos?
The miniMos transposon is used to carry transgenes into the C. elegansgenome by the means of inserting a transgene into a modified Mos1 element (miniMos). The miniMos transposon can carry the inserted transgene (together with a selection marker) into the genome where insertions are generated directly by injecting worms. It will take approximately 1 week to get the result. The insertion site is random and can be identified by inverse PCR on the strain.
The miniMos system contains 4 vectors——transposon with target gene, transposase, mcherry and peer-1 markers. Transposon needs transposase to cut and paste in the genome. mcherry and peer-1 are two negative selecter that can help to select the target strains.
The method has some advantages:
1)The insertion frequency and fidelity is high.
2)The exact insertion site can be determined.
3)Large transgenes can be inserted.
4)The miniMos element is active in C. elegans.
5)Transgenes are expressed in the germline at high frequency.
How to get the worms?
As a classic model organism, C. elegans has many advantages.It is one of the simplest organisms with a nervous system. Research has explored the neural and molecular mechanisms that control several behaviors of C. elegans, including chemotaxis, thermotaxis, mechanotransduction, learning, memory, and mating behaviour. Therefore, C. elegans is a suitable model organism which we can easily manipulate and observe. In our program, we aim to change the physical behaviour of C. elegans by using the two specific optogenetic traits. Through microinjection and selection, we are able to get two strains with two phenotypes of the preference to blue light and the aversion to red light, and the next step is to obtain the single worm with the combination of these 2 different traits.
To achive our objectives, we choose a mating method, because it is less time-consuming and easier to operate than microinjecting the other array to the existed strain.
In general, mating is the pairing of either opposite-sex or hermaphroditic organisms. However, for C. elegans, mating can hybridize two different traits on the next generation by the paring of the hermaphrodite and the male. Besides, one of the greatest advantages of C. elegans is that it can either autocopulation or hybridization, which means that after hybridization, the hermaphrodite can self-fertilize and guarantee to produce a large number of offsprings with identical genetic characteristics.
Here is the mating procedures: Mating procedures: 1.Heat shock to get males(30℃,8h). 2.Parents mating to get the first filial hybrid generation(：=3:4). 3.Single out each hermaphrodite in a small plate. 4.Test the fluorescence of F1. 5.Single out F1 and self-fertilizing to get F2. 6.Test the fluorescence of F2.
References
