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Visuble self-sucking extrinsic gene induction of insect egg

An exogenous gene, self-priming technology, applied to other methods of inserting foreign genetic materials, using vectors to introduce foreign genetic materials, recombinant DNA technology, etc., can solve problems such as expensive and precise instruments and equipment

Inactive Publication Date: 2005-07-27
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This introduction method requires expensive precision instruments and highly skilled experimental operators

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The silkworm eggs of Nistari silkworm variety within 8 hours after laying eggs were used as experimental materials, and the eggs were soaked in 4% formaldehyde for 20 minutes to disinfect the surface of the eggs. The GFP (green fluorescent protein) exogenous gene integrated in the piggyBac transposon plasmid, specifically the pPIGA3GFP plasmid and its auxiliary plasmid pHA3PIG system, is used to introduce the GFP (green fluorescent protein) exogenous gene. The magnification of the solid microscope is 32 times. The DNA plasmid containing the exogenous gene GFP and its auxiliary plasmid are dissolved with ultrapure water, and the DNA concentration is adjusted to 500ng / μl. The amount of the plasmid solution containing the exogenous gene smeared on the surface of silkworm eggs is 50nl. The site where the exogenous gene is smeared is the ventral surface of silkworm eggs. The needle tip diameter of the metal needle piercing the silkworm eggs is less than 0.1mm. The number of...

Embodiment 2

[0023] The silkworm eggs of Viet Nam Bombyx mori within 2 hours after laying eggs were used as experimental materials, and the silkworm eggs were sprayed with 75% alcohol to disinfect the surface of the eggs. The DsRed_ (red fluorescent protein) exogenous gene integrated in the piggyBac transposon plasmid, specifically the pBac[3×P3-EGFP-P9DsRed-LITR] plasmid and its auxiliary plasmid pHA3PIG system, was introduced into the DsRed_ (red fluorescent protein) protein) exogenous gene. The magnification of the solid microscope is 25 times. The DNA plasmid containing the exogenous gene DsRed and its auxiliary plasmid are dissolved with phosphate buffer (pH7.6), and the DNA concentration is adjusted to 200ng / μl. The plasmid containing the exogenous gene smeared on the surface of silkworm eggs The solution volume is 30nl, and the site where the exogenous gene is applied is the dorsal side of silkworm eggs. The needle tip diameter of the metal needle piercing the silkworm eggs is less...

Embodiment 3

[0025] The silkworm eggs of Jiaqiu silkworm variety within 4 hours after laying eggs were used as experimental materials, and the silkworm eggs were soaked in 2% formaldehyde for 30 minutes to disinfect the surface of the eggs. Straight-chain gene is adopted, and the CAT gene with A3 promoter is an exogenous gene. The magnification of the solid microscope is 40 times. The amount of the solution containing the exogenous gene applied on the surface is 100 nl, and the exogenous gene is applied on the surface of silkworm eggs. The needle tip diameter of the metal needle piercing the silkworm eggs is less than 0.01mm. The number of puncture silkworm eggs was 2 times, and the puncture port was not sealed with paraffin after puncture. A total of 5,000 silkworm eggs were punctured in the experiment, and 165 silkworm eggs expressed the CAT gene.

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Abstract

Visual self-sucking method for inducing external gene into insect eggs is carried out by: under telescope, coating solution with external genes on insect eggs, pricking eggs to make external gene into eggs, making external genes combined with insect's gene to form transgenic insects. It makes technicians to do researches on transgenic insects with cheap instrument.

Description

technical field [0001] The invention relates to a visual self-absorption type exogenous gene introduction method of insect eggs. Background technique [0002] The cloned exogenous gene is artificially integrated into the insect genome through biotechnology, and is stably inherited and expressed. Such insects are called transgenic insects. Transgenic insects can form a new species with corresponding exogenous genes, and their utilization value lies in: [0003] (1) Carry out basic theoretical research; [0004] (2) Production of exogenous proteins; [0005] (3) Molecular breeding of commercial insects; [0006] (4) Carry out biological control of pests. [0007] When conducting research on transgenic insects, the cloned exogenous gene is generally introduced into the egg stage by microinjection. This introduction method requires expensive precision instruments and highly skilled experimental operators. Contents of the invention [0008] The purpose of the present inve...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/87
Inventor 钟伯雄
Owner ZHEJIANG UNIV
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