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Algae feed capable of preventing avian influenza and application thereof

A technology of avian influenza and Chlamydomonas lychee, applied in the field of algae feed, can solve the problem of the absence of avian influenza antigen genes

Active Publication Date: 2014-05-28
深圳市微宇藻业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, there is no technical solution or related reports about using Chlamydomonas reinhardtii as a host to express the avian influenza antigen gene

Method used

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  • Algae feed capable of preventing avian influenza and application thereof
  • Algae feed capable of preventing avian influenza and application thereof
  • Algae feed capable of preventing avian influenza and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] [Example 1] Optimization and amplification of avian influenza antigen genes

[0060] According to the known avian influenza antigen genes (GenBank accession number: AY609312) and the codon preference of Chlamydomonas reinhardtii, the codons that can be preferentially expressed in the nucleus of Chlamydomonas reinhardtii are selected to replace the known avian influenza antigen genes , and the amino acid sequence remains unchanged, thereby obtaining the HAV1 and HAV1' genes suitable for the nuclear genetic expression of Chlamydomonas reinhardtii, as shown in SEQ ID NO.1 and SEQ ID NO.2 respectively; The HAV2 and HAV2' genes are shown as SEQ ID NO.3 and SEQ ID NO.4 respectively; HAV3 and HAV3' genes suitable for the expression of Chlamydomonas reinhardtii chloroplasts are optimized, as shown in SEQ ID NO.5 and SEQ ID NO. .6 shown. The above codon-optimized avian influenza antigen genes SEQ ID NO.1-SEQ ID NO.6 were artificially synthesized by Sangon Bioengineering (Shangh...

Embodiment 2

[0075] [Example 2] Selection and cultivation of transgenic recipient algal strains

[0076] The selected transgenic recipient algal strains in the present invention are: cell wall-deficient Chlamydomonas reinhardtii (Chlamydomonas reinhardtii, cc-849, purchased from the Chlamydomonas Genetics Center of Duck University, USA) and cytochrome b-deficient Chlamydomonas reinhardtii (cc-2654, purchased from the United States) from the Chlamydomonas Genetics Center at Duck University, USA). TAP was selected as the medium for the transgenic algal strain, and its preparation method was as follows: 25mL of 4×Beijerinck salts (4g MgSO 4 .7H 2 O, 2gCaCl 2 .2H 2 O, 16g NH 4 Dissolve Cl in deionized water, make up to 1L), 2.42g Tris, 1mL 1M(K)PO 4 , 1mL Trace trace element mixture (50g Na 2 EDTA, 1.1g (NH 4 ) 6 Mo 7 O 24 , 1.57gCuSO 4 .5H 2 O, 1.61g CoCl 2 .6H 2 O, 4.99g FeSO 4 .7H 2 O, 22g ZnSO 4 .7H 2 O, 5.6gMnCl 2 .4H 2 O, 11.4g H 3 BO 3 Dilute to 1L deionized water)...

Embodiment 3

[0077] [Example 3] Construction of a genetic expression vector for transgenic avian influenza antigen gene Chlamydomonas reinhardtii

[0078] (1) Construction of avian influenza antigen gene Chlamydomonas reinhardtii nuclear genetic expression vector

[0079] It can be seen from Example 1 that the plasmid pUC57HAV1 contains the HAV1 gene, and pUC57HAV1' contains the HAV1' gene. Since the appropriate restriction enzyme sites were added while optimizing the sequence, the restriction enzymes Nhe I and Pmac Ⅰ Double enzyme digestion (purchased from Fermentas) to obtain HAV1 and HAV1' genes. Plasmid pH105 (Wu JX et al. Efficient expression of green fluorescent protein (GFP) mediated by a chimeric promoter in Chlamydomonas reinhardtii. Chinese Journal of Oceanology and Limnology. Vol.26No.3, P.242-247, 2008 was digested with EcoRⅠ ) to obtain a portion of about 731 bp containing the HSP70A-RBCS2 promoter and RBCS2 terminator, which was inserted into pSP124 (Lambreras v, Stevens DR,...

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Abstract

The invention discloses an algae feed capable of preventing avian influenza and application thereof, discloses an artificially synthesized avian influenza antigen gene optimized by a codon preferred by chlamydomonas reinhardtii, establishes a recombined expression vector suitable for expressing the avian influenza antigen gene in the chlamydomonas reinhardtii, and transfers the recombined expression vector into host chlamydomonas reinhardtii to obtain transgenic chlamydomonas reinhardtii. Animal experiments show that a mouse eating the transgenic chlamydomonas reinhardtii can generate an avian influenza antibody; the algae feed has a wide application prospect and can prevent the avian influenza.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an algae feed capable of preventing bird flu, a preparation method and application thereof. Background technique [0002] In recent years, relevant data and facts show that for Asia alone, a relatively mild outbreak of avian flu will cost 90-110 billion US dollars. If it is a global outbreak of bird flu, it may cause a global economic recession. At the same time, according to economists' estimates, at the time of the outbreak of bird flu, the direct economic losses caused by bird flu to trade and tourism in the affected areas are also incalculable. From the four worldwide influenza outbreaks in the last century, three have caused great harm to human beings, namely, the spread of influenza A virus that broke out in Spain in 1918, and the outbreak of A2 (H2N2) in 1957. The spread of subtype influenza virus and the outbreak of A3 (H3N2) subtype influenza in 1968 caused soc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/82C12N1/13A61K39/145A61P31/16C12R1/89
Inventor 胡章立李建成
Owner 深圳市微宇藻业科技有限公司