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Studies on the Effect of Components of Artificial Diet on Gene Expression Efficiency and Recombinant Virus Peroral Infection of rBmNPV-Bm System

Author: ShangJinYan
Tutor: CuiWeiZheng;ZhangZhiFang
School: Shandong Agricultural University
Course: Special Economic Animal Feeding
Keywords: Silkworm Baculovirus expression vector system Expression levels Food under infection Artificial diet
CLC: Q789
Type: Master's thesis
Year: 2003
Downloads: 22
Quote: 1
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Abstract


The BEVS is one of the four expression systems development and use in the field of genetic engineering, production of medical vaccines and protein expression systems rBmNPV-Bm most promising, but in practical there are still some issues to be resolved, such as the use of the expression system drugs using sterile silkworm, requires the use of sterile artificial diet rearing; Second, the recombinant virus pathogenicity weak conventional mulberry leaves fertile conditions generally can not eat under infection can only take a single head microinjection method vaccination virus, cumbersome operation, ergonomics is low, it is difficult to meet the requirements of large-scale production; addition, the amount of exogenous gene expression by a variety of factors to be further enhanced expression activity. For this reason, the study is based on artificial diet sericulture, to increase the amount of exogenous gene expression and recombinant virus infection rate was food under and in animal husbandry value phytase gene as a reporter gene, fresh artificial diet composed of exogenous gene expression rBmNPV-Bm system activity, and recombinant virus infection techniques, and obtained the following results. Workers feed mulberry leaf powder, soy flour, water, vitamins, inorganic salts and other nutrients content, activity of in vivo expression of foreign genes in silkworm have a significant impact, the mulberry leaf powder content of 10%, the content of soy flour 35%, moisture dosage dry weight of 1.9 times the amount of vitamin C, 1%, the amount of vitamin B mixture 0.25% VB6 2mg/100g dry matter, inorganic salts add the amount of 1% of the phytase gene expression the highest activity; phytase activity between the highest and lowest area, respectively, a difference of 1.92 times, 1.57 times, 3.23 times and 2.13 times, 2.35 times, 4.54 times and 1.78 times respectively. 2. Feed composition of the phytase gene expression activity influence and impact on the the silkworm body weight growth is basically the same, that is, to express the high activity of the feed zone, the silkworm weight. 3. The five instar silkworm After the impact of low temperature (5 ℃ 24h) or high temperature (38 ℃ 10h), the food of recombinant virus infection rates were increased, the highest infection rates were 83% and 679/0, without the high and low temperature The control area of ??the impact of the incidence rate of O. After the impact of the low and high temperature, the pH value of the silkworm intestinal fluid were significantly decreased. 4. To explore the effect of fluorescent whitening agent VBL and Tinapal LPW food to improve recombinant virus infection rates to identify the VBL Tim food concentration and licking infection rates under food. The results show that both have the duty to promote the role of infection in the recombinant virus food, but the the VBL synergies than Tinapal LPW and less impact on the growth and development of silkworm. Fifth instar silkworm the 1% VBL24h before Tim food from the recombinant virus and 1% VBL mixed Tim Fresh, a sense of lt; WP = gt; the dyeing rate may reach 90%. 5. Silkworm Tim food VBL, resulting in intestinal fluid pH decreases intestinal fluid protease activity, the extent of its influence to increase food concentration increased with Tim, which Tim Fresh 2% VBL 24h intestinal fluid protease activity decreased to 4.31 times. Therefore, The fluorescent brighteners can lead to abnormal the digestive physiological function of short-term destruction of the normal function of the defense, so that the recombinant extracellular appearance virus through the midgut to reach the blood chamber, triggering infection.

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering) > Application of genetic engineering
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