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A recombinant Hansenula polymorpha expressing Zika virus structural protein e and its construction method

A technology of Hansenula polymorpha and structural protein is applied in the field of construction of Hansenula polymorpha expression system, which can solve the problems of low expression efficiency and non-expression of ZKE protein, and achieve low cost, good immunogenicity and production potential. big effect

Active Publication Date: 2020-10-13
NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION +1
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  • Application Information

AI Technical Summary

Problems solved by technology

The main difficulty may lie in: affected by the structure of the ZKE protein, the expression efficiency of the ZKE protein in the host cell is low or even not expressed

Method used

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  • A recombinant Hansenula polymorpha expressing Zika virus structural protein e and its construction method
  • A recombinant Hansenula polymorpha expressing Zika virus structural protein e and its construction method
  • A recombinant Hansenula polymorpha expressing Zika virus structural protein e and its construction method

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Experimental program
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Embodiment Construction

[0021] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0022] (1) Construction of pHMOXG-alpha-ZKE recombinant expression vector containing Zika virus structural protein E gene fragment 1. According to the reported Zika virus structural protein E gene information (GenBank: KU820899.2), artificially design Zika virus to be expressed Viral structural protein E gene fragment (designed in May 2017), the two ends of which can be added with amplification primers (primer 1 and primer 2, designed in July 2017) EcoRI (carbon terminus, see sequence underlined) position) and restriction site NotI (nitrogen terminus, see sequence italicized position). The artificially designed Zika virus structural protein E gene fragment sequence is shown in SEQ.ID.NO.1, which is an artificially synthesized sequence.

[0023] 2. After the plasmid (pMD18-T, Takara) containing the designed Zika virus structural protein E gene...

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Abstract

The invention discloses recombinant hansenula polymorpha for expressing Zika virus structural protein E and a construction method thereof. According to the recombinant hansenula polymorpha, a gene segment of the artificially synthesized Zika virus structural protein E is introduced into hansenula polymorpha to obtain a recombinant bacterium for expressing the Zika virus structural protein E. A nucleotide sequence of the gene segment of the Zika virus structural protein E is shown as SEQ ID NO.1. The recombinant hansenula polymorpha can be used for producing the recombinant Zika virus structural protein E and has the advantages of simple purification steps, low cost and great industrial production potential.

Description

technical field [0001] The invention belongs to the technical field of genetic vaccine engineering and relates to the construction of a Hansenula polymorpha expression system for Zika virus glycoprotein. Background technique [0002] Zika virus belongs to Flaviviridae family (Flaviviridae), which is similar to dengue virus, yellow fever virus, Japanese encephalitis virus and West Nile virus. Zika virus is a single-stranded positive-sense RNA virus with a long genome. About 10.8kb, containing a single open reading frame, the viral protein is composed of a single polyprotein precursor, which is digested by host protease and viral protease, including 3 structural proteins (C, prM / M, E) and 7 nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5). Similar to other flaviviruses, the structural protein E (ZKE) of Zika virus is highly conserved and contains multiple neutralizing epitopes. In addition, the ZKE protein is the main protein that binds to the cell membrane rec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/81C12N1/19C07K14/18C12R1/78
CPCC07K14/005C12N15/815C12N2770/24122
Inventor 李永东钱卫东林文凭焦素黎刘昱慧
Owner NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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