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Duck Tembusu virus e protein truncated gene, recombinant duck plague virus and its construction method and application

A technology of duck Tembusu virus and duck plague virus, which can be applied in the direction of virus/phage, virus, application, etc., can solve the problem of low expression of E protein

Active Publication Date: 2020-07-07
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims at the problem of low expression of Duck Tembusu virus E protein of recombinant duck plague virus in the prior art, and provides a duck Tembusu virus E protein truncated gene, which is inserted into the recombinant duck After the recombinant duck plague virus obtained from the genome of pestivirus infects chicken embryo fibroblasts (CEFs), the protein expression of duck Tambusu virus E protein truncated body increases

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  • Duck Tembusu virus e protein truncated gene, recombinant duck plague virus and its construction method and application
  • Duck Tembusu virus e protein truncated gene, recombinant duck plague virus and its construction method and application
  • Duck Tembusu virus e protein truncated gene, recombinant duck plague virus and its construction method and application

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Embodiment 1

[0037] 1. Sequence optimization, primer design and synthesis

[0038] The E gene (sequence reference GenBankJF270480.1) was optimized and synthesized by GenScript Company with reference to chicken and duck codon bias, and the original E gene (sequence shown in SEQ ID NO.1) was obtained, which was codon-optimized using chicken as a host. E gene (sequence shown in SEQ ID NO.2), codon-optimized E gene (sequence shown in SEQ ID NO.3) using duck as host. The primers were designed with reference to the sequence GenBank (EU082088.2) and the optimized sequence of the Duck Tembusu virus E gene, and were synthesized by Shanghai Sangong.

[0039] The primers used in this paper are listed in Table 1:

[0040] Eori+ / Eori- primer pair is used to amplify the original E gene (Eori);

[0041] Eori+ / E451(ori-) is used to amplify the C-terminal truncated E gene (reserve 1-451 aa) (E451-ori);

[0042] Etpa(ori+) / Eori-used to amplify the original E gene (Etpa-ori) with the chicken TPA signal pe...

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Abstract

The invention discloses a duck Tembusu virus E protein truncated gene, a recombinant duck plague virus, a construction method and an application. The nucleotide sequence of the duck Tembusu virus E protein truncated gene is shown in SEQ ID No.4. A recombinant duck plague virus, wherein the duck Tembusu virus E protein truncated gene is inserted into the genome of the recombinant duck plague virus. The present invention screens and obtains a truncated gene of duck Tembusu virus E protein, inserts the E protein truncated gene into the genome of recombinant duck plague virus to obtain a new recombinant duck plague virus, and infects CEFs After cells, the E protein truncated gene is highly expressed. The recombinant duck plague virus inserted with the E protein truncated gene can be used for the preparation of duck plague virus and duck Tembusu virus dual vaccine.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a truncated gene of duck Tembusu virus E protein, a recombinant duck plague virus, a construction method and an application. Background technique [0002] Since April 2010, a new infectious disease has broken out in laying ducks and breeding ducks in the main duck production areas of my country. The main clinical features of the disease are rapid spread, sudden drop in egg production, and hemorrhagic oophoritis. . Through pathogen isolation and identification and full sequence determination, it is clear that the pathogen belongs to Flaviviridae, Flavivirus, Mosquito-borne, and Ntaya virus groups, and has similarities with this group of Tembusu viruses (TMUV). The recent genetic evolution relationship can be considered as a new member of TMUV, tentatively named Duck Tembusu virus (DTMUV) (Su J, Li S, Hu X, et al. Duck egg-drop syndrome caused by BYD virus, a new Tembusu-related flavi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/40C12N7/01A61K39/245A61K39/12A61P31/22A61P31/14
CPCA61K39/00C07K14/005C12N7/00C12N2710/16021C12N2710/16034C12N2770/24122C12N2770/24134Y02A50/30
Inventor 陈柳张存余斌华炯钢倪征叶伟成云涛
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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