Novel method for infecting silkworms with recombinant baculovirus
A technology of virus infection and silkworm baculovirus, which is applied in animal husbandry and other fields, can solve the problems of virus liquid flowing out of the body, cumbersome operation, and long time-consuming, so as to simplify the virus infection method, solve the cumbersome operation, and improve work efficiency.
Inactive Publication Date: 2013-01-23
ZHEJIANG UNIV
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Problems solved by technology
However, the current infection technology is to use the method of directly injecting the recombinant virus into the silkworm for infection and expression. In the actual operation process, it needs to be injected one by one silkworm, which takes a long time and is cumbersome to operate. Due to the peristalsis of worms and other reasons, the virus liquid flows out of the body, which eventually leads to the failure of infection. Therefore, the current traditional percutaneous injection method has obvious defects and low work efficiency.
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[0012] Introduce the inventive method in further detail below:
[0013] 1. Materials: Recombinant silkworm baculovirus was constructed using a very mature technology. For specific methods, please refer to papers published at home and abroad, such as "Using Bacterial Transposons to Construct Bac- to-Bac Rapid Gene Expression System", 32(2): 183-188. A recombinant silkworm baculovirus expressing the reporter gene green fluorescent protein EGFP is constructed by using the above-mentioned Bac-to-Bac technology of the silkworm, and the virus is used to transdermally infect the silkworm larvae using the method of the present invention.
[0014] In the following examples, a silkworm hybrid species named Qiufeng×Baiyu was used, and silkworm larvae were fed with mulberry leaves at a temperature of 23-25°C.
[0015] In the following examples, the method for percutaneously infecting silkworms with recombinant baculovirus is as follows:
[0016] Concentrate the 3rd or 4th instar la...
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The invention discloses a novel method for infecting silkworms with recombinant baculovirus. The method comprises the following steps: silkworm larvae are gathered into a cultivation frame; in a standard that each 100 silkworm larvae are ejected with 5ml of recombinant baculovirus liquid, the recombinant baculovirus liquid with 106pfu is ejected to the body surfaces of the silkworm larvas; and the silkworm larvae are not fed with mulberry leaves, and after most part of the baculovirus liquid is dried, the silkworm larvae are fed in an ordinary method till the silkworm larvae have the symptom of virus infection. The method disclosed by the invention has the advantages that a traditional inoculation method of percutaneous infection which is used directly is changed, the inoculation operation is simplified, the inoculation time is saved, and the working efficiency of an insect baculovirus expression system is improved.
Description
technical field [0001] The invention relates to a recombinant virus infection technology, in particular to a method for infecting silkworms with a recombinant virus. Background technique [0002] BacuLovirus Expression Vector System (BEVS for short) is a recombinant protein production system that uses recombinant insect baculovirus carrying an exogenous target gene as a vector to express and produce in insects or insect cultured cells. Because the system has the advantages of large-scale high-efficiency expression production using individual insects or their cultured cells, it has become one of the most effective eukaryotic expression systems at present. At present, BEVS mainly includes Autographa californica nuclear polyhedrosis virus ( Autographa californica nucleopolyhedrosis virus (AcNPV for short) and the silkworm nuclear polyhedrosis virus (AcNPV), which is widely used in China and Japan Bombyx mori nucleopolyhedrosis virus, BmNPV for short) two expression systems ...
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IPC IPC(8): A01K67/04
Inventor 杨锐沈运旺吴小锋
Owner ZHEJIANG UNIV