Method for cloning male genomes of lepidopterous insects

A lepidopteran insect and genome technology is applied in the field of insect bioengineering to solve the mating problem and achieve the effect of low cost

Inactive Publication Date: 2010-09-15
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, among the Lepidoptera insects, only the silkworm has androgenic technology, but this

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The male genome clone of the castor silkworm (Philosamia cynthiaricini Boisduval) yellow blood white skin system Gaozhou No. 1 variety was obtained from the eggs of the Bombyx mori Dazao variety of Lepidoptera. Variety No. 1 is preserved and provided by the Sericulture Research Institute of Soochow University. The specific operation steps are as follows:

[0042] (1) The recipient silkworm (Bombyx mori Dazai variety) is raised at an ambient temperature of 25°C, and the female pupa is also protected at 25°C until eclosion; the female pupa receives 70krad of Co-60γ-ray radiation 2d to 3d before eclosion to destroy the oocyte nucleus;

[0043] (2) Donor castor silkworm (Castor silkworm Gaozhou No. 1) larvae were reared at 25°C, and 2d to 3d after pupation, when the pupal skin was just colored, they were transferred to an environment of 5°C to 8°C and a relative humidity of 75% to 80%. Adjust the protection, take out a certain number of male and female pupae every day and p...

Embodiment 2

[0051] The male genome clone of Bombyx mandarina L. was obtained from the eggs of Bombyx mandarina L., which was preserved and provided by the Institute of Sericulture of Soochow University. The wild silkworm was obtained from The mulberry garden of Soochow University collects it by itself and then feeds and purifies it. The specific operation steps are as follows:

[0052] (1) The recipient wild silkworm was reared at 27°C, and the female pupa was also protected at 26°C until eclosion; the female pupa received 70krad of Co-60γ-ray radiation 2d to 3d before eclosion;

[0053] (2) Donor silkworm larvae are reared at 25°C, and after pupation, they are adjusted and protected in an environment of 23°C to 26°C and a relative humidity of 75% to 80%, so as to ensure that there are eclosions from 2 days before the eclosion of the wild silkworm female moth to the day of eclosion Silkworm female and male moths;

[0054] (3) At an ambient temperature of 25°C, the male and female moths o...

Embodiment 3

[0060] The male genome clone of the green tussah silkworm (Anfheraepernyi Guerin-Meneville) was obtained from the eggs of the silkworm Dazao variety of Lepidoptera Anfheraepernyi Guerin-Meneville. provided, the specific steps are as follows:

[0061] (1) Recipient silkworms were reared at an ambient temperature of 25°C, and the female pupae were also protected at 25°C until eclosion; the female pupae received 70krad of Co-60γ-ray radiation 2d to 3d before eclosion;

[0062] (2) The larvae of the donor tussah silkworm (green variety of tussah silkworm) are raised at 20°C to 24°C, and the diapause pupae are adjusted and protected at 8°C to 12°C and a relative humidity of 75% to 80%; Protect at 25°C to ensure that the male tussah moths emerge from 2 days before the emergence of the female silkworm moths to the day of emergence;

[0063] (3) The seminal vesicles of tussah silkworm male moths were dissected under sterile conditions, washed 3 times with sterile saline, and the seme...

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PUM

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Abstract

The invention discloses a method for cloning male genomes of lepidopterous insects, which comprises the following steps: (1) radiating female pupae of receptor insects in 2d-3d before eclosion to destroy oocyte nuclei; (2) extracting semen of male moths of donor insects; (3) carrying out artificial insemination on virgin moths of the receptor insects; (4) collecting ova propagated by female moths after artificial insemination treatment; (5) carrying out androgenesis treatment on the propagated ova obtained in the step (4) to obtain androgenesis ova; and (6) screening and treating the androgenesis insect ova obtained in the step (5), and carrying out hatching treatment to obtain G1-generation male genome clones of the donor insects. The method of the invention can avoid injury of nucleus transplantation required for a nucleus transplantation cloning technology and an artificial reproduction method, fusion treatment of sperm nuclei and ovoplasm and genome activation stimulation, has low aberration rate of cloned posterity, can obtain individual posterity, tissues and cells which are highly consistent with sperm donor insects, and can obtain high-quality materials for evaluating influence of ovoplasm to cloned animals.

Description

technical field [0001] The invention belongs to the field of insect bioengineering, and in particular relates to a method for cloning the male genome of lepidopteran insects. Background technique [0002] The biological meaning of the word clone refers to the asexual reproduction line as a noun, and refers to the process of asexual reproduction as a verb. As a noun, cloning has three levels of meanings: cloning organisms, cloning cells and cloning genes. A cloned organism refers to a multicellular organism that is genetically the same as another (group) organism; a cloned cell refers to a pure cell line formed by the division and reproduction of the same ancestor cell; a cloned gene refers to a certain gene (or fragment) of an organism Copy product. Cloning technology is a commonly used research and application technology in the field of life sciences. It is mainly used for reproductive cloning to produce offspring organisms and research cloning or medical cloning to produ...

Claims

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Application Information

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IPC IPC(8): A01K67/033
Inventor 徐世清袁红霞司马杨虎甘丽萍季明明
Owner SUZHOU UNIV
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