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Construction and culture methods of recombinant rabies virus

A technology of rabies virus and culture method, which is applied in the field of construction of recombinant rabies virus expressing 3G gene, can solve problems such as potential safety hazards, and achieve the effects of wide applicability, high rescue efficiency, and excellent immunogenicity

Inactive Publication Date: 2015-05-27
山东华宏生物工程有限公司
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Problems solved by technology

In view of the large safety hazards in the use of attenuated live vaccines, the Ministry of Agriculture of my country stipulates that inactivated vaccines must be used for the prevention of rabies

Method used

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  • Construction and culture methods of recombinant rabies virus
  • Construction and culture methods of recombinant rabies virus
  • Construction and culture methods of recombinant rabies virus

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Experimental program
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Embodiment 1

[0043] 1. Preparation before construction

[0044] 1A) Construction of cDNA

[0045] According to the SAD Bern (Sanafox) whole genome sequence published by Gene Bank, the SAD Bern (Sanafox) strain genome sequence is divided into 8 independent genome fragments (such as figure 1 shown), using the base sequences of the above 8 independent gene fragments as templates, using reverse transcriptase to carry out bio-synthesis (Nanjing Zhongding Biotechnology Co., Ltd.), to obtain the same complementary DNA as the 8 independent genome fragments ( cDNA) fragments F1~F8, and introduced PmeI restriction site and hammerhead ribozyme (HamRz) at the 5' end of F1, and introduced NotI restriction site and hepatitis D virus ribozyme at the 3' end of F8, thereby The F1 gene fragment containing the hammerhead ribozyme and the F8 gene fragment containing the hepatitis D virus ribozyme are obtained. Then clone the synthetically prepared gene fragments of F1~F8 into the pCR-Blunt vector plasmid re...

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Abstract

The invention provides a recombinant rabies virus strain. A nucleotide sequence as shown in SEQ ID NO.1 is formed by inserting two rabies virus G genes into a rabies virus genome which is obtained by reverse genetic operation of a whole genome sequence of a SAD Bern rabies virus. The constructed recombinant rabies virus has better security and immunogenicity and is suitable for preparing an inactivated rabies vaccine.

Description

technical field [0001] The present invention belongs to the field of recombinant vaccines. In particular, it relates to the construction of a recombinant rabies virus expressing 3G gene. Background technique [0002] Rabies is an important zoonotic disease, causing approximately 60,000 deaths per year. In many contexts, dogs are still the main cause of rabies in humans, and in developed countries, rabies virus is mainly found in wild animal hosts. Practice has proved that immunizing animals such as dogs can significantly reduce the incidence of human rabies, so immunizing animals is very important for the control of rabies. [0003] Rabies virus belongs to the genus Lyssavirus in Rhabdoviridae. The genome is a single-stranded negative-sense RNA without segments, with a total length of about 12Kb, encoding five viral proteins: nucleoprotein N, phosphoprotein P, matrix protein M, and glycoprotein G and RNA polymerase L. Glycoprotein G, located on the capsule, is the main v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12R1/93
Inventor 刘澜朱万光李城
Owner 山东华宏生物工程有限公司
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