Method for producing cultivated silkworm novel green cocoon practical variety by using GFP protein
A silkworm and variety technology, applied in the fields of bioengineering and biotechnology, can solve problems such as no research reports, and achieve the effect of simplifying the spinning process, unique optical characteristics, and wide use.
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Embodiment 1
[0028] The first step: use the practical silkworm variety "Bibo" (the original species of Japanese white cocoon) as the raw material, remove the diapause of the first-laid eggs, and use an Eppendorf microinjector to inject 10- 15n1, a total concentration of 400ng / u1 recombinant piggyBac vector and helper vector with red fluorescent protein tagged by nerve and eye-specific promoters and green fluorescent protein (GFP protein) promoted by silk fibroin heavy chain promoter, co-injected into 1500 In the silkworm eggs, the injection port was sealed with non-toxic glue, and after being sterilized in 35% formaldehyde vapor for 5 minutes, they were placed in a high-humidity environment at 25°C and a relative humidity of more than 85% to accelerate the hatching, and the hatched 73 heads of G0 The generation ant silkworms were collected and raised with artificial feed until they became moths, and 40 G0 generation silkworm moths were obtained. A total of 27 G1 generation silkworm eggs wer...
Embodiment 2
[0032] The first step: using the practical silkworm variety "872" (the original species of Japanese white cocoon) as the raw material, the newly laid eggs were released from diapause, and 3-6 hours after laying, the 10- 15nl, total concentration of 400ng / ul recombinant piggyBac vector and helper vector with red fluorescent protein tagged by nerve and eye-specific promoter and green fluorescent protein (GFP protein) promoted by silk fibroin heavy chain promoter, co-injected into 2500 In the silkworm eggs, seal the injection port with non-toxic glue, sterilize in 35% formaldehyde vapor for 5 minutes, and place them in a high-humidity environment at 25°C and a relative humidity above 85% to accelerate the hatching. The hatched 163 G0 The generation ant silkworms were collected and raised with artificial feed until they became moths, and 121 G0 generation silkworm moths were obtained. A total of 63 G1 generation silkworm eggs were obtained through selfing or backcrossing, and 63 G1...
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