Trivalent HA-2HA1-LTB fusion expression vector of lotus corniculatus specifically-expressed H5N1 antigen protein

An antigen protein, fusion expression technology, applied in the field of genetic engineering, can solve the problems of traditional vaccine development and difficult immunization routes

Inactive Publication Date: 2014-06-18
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The continuous mutation of H5N1 highly pathogenic avian influenza virus and its ability to spread through migratory birds have brought great difficulties to the development of traditional vaccines and immunization methods

Method used

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  • Trivalent HA-2HA1-LTB fusion expression vector of lotus corniculatus specifically-expressed H5N1 antigen protein
  • Trivalent HA-2HA1-LTB fusion expression vector of lotus corniculatus specifically-expressed H5N1 antigen protein
  • Trivalent HA-2HA1-LTB fusion expression vector of lotus corniculatus specifically-expressed H5N1 antigen protein

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Experimental program
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Embodiment 1 3

[0062] Example 1 Construction of trivalent complete HA-LTB fusion plant expression vector

[0063] 1. Construction process of plant expression vector

[0064] The codons of HK97HA, HK07HA1, SX06HA1 and immune adjuvant LTB genes were optimized, and the upstream sequences of the start codons of each target gene were modified, and the modified genes were synthesized in full sequence. On the premise of not changing the amino acid composition of avian influenza hemagglutinin antigen protein (HA) and immune adjuvant Escherichia coli heat-sensitive toxin B subunit (LTB), the codons of the antigen protein were optimized to promote the increase of foreign antigen protein in Expression levels in recipient plant japonicus japonicus. The optimized HK97HA, HK07HA1, SX06H1A and immune adjuvant LTB gene sequences are shown in SEQ ID No.2, 4, 6 and 8.

[0065] PCR reaction

[0066] Mar-2E+35S-Ω and poly(A)-Nos-Mar long fragment PCR reaction conditions: pre-denaturation at 94°C for 5 min, t...

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Abstract

The invention relates to the field of genetic engineering and specifically relates to a trivalent HA-LTB fusion expression vector of a lotus corniculatus specifically-expressed H5N1 antigen protein. At a multiple cloning site of a lotus corniculatus plant expression vector, in a direction from a 2E+35 stator to a poly(A)-Nos terminator, an optimized HK97HA gene as shown in a nucleotide sequence SEQ ID No.2, an optimized HK07HA1 gene as shown in a nucleotide sequence SEQ ID No.4, an optimized SX06HA1 gene as shown in a nucleotide sequence SEQ ID No.6 and an optimized immunologic adjuvant LTB gene as shown in a nucleotide sequence SEQ ID No.8 are inserted in the forward direction in sequence. According to the invention, an established foreign protein high-level expression platform is utilized to express, so that a foundation is laid for developing broad-spectrum H5N1 bird flu plant edible vaccines.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a trivalent HA-2HA1-LTB fusion expression vector for japonicus japonicus specifically expressing H5N1 antigenic protein. Background technique [0002] The continuous mutation of H5N1 highly pathogenic avian influenza virus and its ability to spread through migratory birds have brought great difficulties to the development of traditional vaccines and immunization methods. Avian influenza virus has a very high frequency of mutation and recombination, coupled with its high toxicity and potential ability to infect humans, making the prevention and control of avian influenza a worldwide problem. The ever-spreading epidemic has once again sounded the alarm for us. Passive treatment based on the subtype of the diseased virus is often lagging and blind, and the avian influenza virus has the characteristics of host diversification and migration with migratory birds. Only a mature vaccin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84
Inventor 吴燕民何景成张伟伟卢运明袁蓓王维部周美亮郑平唐益雄陈肖英
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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