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Expression quantity of increasing insect rhabdovirus system exogenous gene by utilizing insect hormone

A baculovirus, foreign gene technology, applied in the field of biotechnology genetic engineering, can solve the problem of no host application of insect hormones, etc.

Inactive Publication Date: 2003-04-16
中国农业科学院蚕业研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Applying insect hormones to baculovirus-infected hosts may restore the host's normal physiological balance and improve the efficiency of the protein synthesis system, but there is no specific method of administering insect hormones to baculovirus-infected hosts at home and abroad

Method used

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  • Expression quantity of increasing insect rhabdovirus system exogenous gene by utilizing insect hormone
  • Expression quantity of increasing insect rhabdovirus system exogenous gene by utilizing insect hormone
  • Expression quantity of increasing insect rhabdovirus system exogenous gene by utilizing insect hormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The silkworms of the JY variety were reared to the fifth instar according to conventional methods, and 15 silkworms with the same average body weight were selected as a group 48 hours after feeding, and each silkworm was inoculated with 1.0×10 5 pfu Bm-phy (5μl inoculum containing 2.0×10 7 pfu / ml of Bm-phy and 0.8mg MH / ml), a total of 12 groups. When most of the silkworms infected with Bm-phy became ill and died frequently, blood samples were collected and stored at -20°C to be tested for phytase activity. After the Bombyx mori were treated by mixing MH in the virus solution and injecting them together, the average phytase activity per unit volume of silkworm hemolymph was 59.80% higher than that of the control, and the body weight increased by 5.2% compared with the control. The difference is extremely significant ( image 3 ), indicating that after applying 4 μg of MH, the expression level per unit volume can be significantly increased, and the body weight of each s...

Embodiment 2

[0026] The silkworms of the JY variety were reared to the fifth instar according to conventional methods, and 15 silkworms with the same average body weight were selected as a group 48 hours after feeding, and each silkworm was inoculated with 1.0×10 5 pfu Bm-phy (5μl inoculum containing 2.0×10 7 pfu / ml of Bm-phy and 0.8mg MH / ml), a total of 12 groups. When most of the silkworms infected with Bm-phy became ill and died frequently, blood samples were collected and stored at -20°C to be tested for phytase activity. After the Bombyx mori were treated by mixing MH in the virus solution and injecting them together, the average phytase activity per unit volume of silkworm hemolymph was 59.80% higher than that of the control, and the body weight increased by 5.2% compared with the control. The difference is extremely significant ( image 3 ), indicating that after applying 4 μg of MH, the expression level per unit volume can be significantly increased, and the body weight of each s...

Embodiment 3

[0029] With the JY1 variety as the material, the 5th instar was raised normally and injected with 1.0×10 5 pfu of Bm-phy recombinant virus, and 2 μg, 4 μg, 6 μg / head of MH were added to Bm-phy respectively. 3×15 silkworms with similar average body weight were used as a group of experimental materials, and the experiment was repeated three times. Blood samples were collected and stored at -20°C, and phytase activity was determined. The results are as follows: the activity of phytase per unit volume injected with 2μg, 4μg, and 6μg MH to each silkworm was significantly higher than that of the control. Among them, the effect of injecting 4 μg MH is the best, and the activity of hemolymph phytase per unit volume is increased by 52.2% compared with the control; the effect of injecting 6 μg MH per silkworm is the second, which is 46.7% higher than the control; Poor, only increased by 36.6% compared with the control. The body weight of diseased silkworms injected with different amo...

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Abstract

A process for increasing the expression to exogenous gene in the insect baculovirus system by insect hormones features that the molting hormone (MH), the juvenile hormone (JH) and their mixture are used to increase the expression efficiency of exogenous gene and polyhedron protein gene in the insect baculovirus system and the copying efficiency of insect baculovirus in insect cells.

Description

1. Technical field [0001] The invention relates to the field of biotechnology genetic engineering, in particular to the application of insect hormones to increase the expression of exogenous genes in insect baculovirus systems. 2. Background technology [0002] The insect baculovirus expression system of present application mainly contains two, and one is to use Autographa Californica Nuclear Polyhedrosis Virus (AcMNPV) as a carrier, with Fall Armyworm (Spodoptera frugiperda, Sf) cells, Cabbage caterpillar (Trichopiusta ni, Ai-5) cells or insects as the host; the other uses Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) as the vector, expressed in silkworm cells or silkworm worms and pupae Host. Both expression systems use strong promoters such as polyhedrin gene or P10 gene or very early gene IE-1 of their respective viral vectors as regulatory elements for exogenous gene expression, by placing the exogenous target gene under the control of a strong promoter The recombina...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/67C12N15/866
Inventor 张志芳何家禄王厚伟肖庆利李卫国周亚竟
Owner 中国农业科学院蚕业研究所
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