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A kind of algae feed that can prevent bird flu and its application

A bird flu and Rhine coat technology, applied in the field of algae feed, can solve the problem of the absence of avian flu antigen genes and other problems

Active Publication Date: 2016-05-25
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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  • A kind of algae feed that can prevent bird flu and its application
  • A kind of algae feed that can prevent bird flu and its application
  • A kind of algae feed that can prevent bird flu and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

Embodiment 2

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

[0076] The transgenic recipient algal strains selected by the present invention are: cell wall defect type Chlamydomonas reinhardtii (Chlamydomonas reinhardtii, cc-849, purchased from the Chlamydomonas Genetic Center of Duck University in the United States) and cytochrome b-defective type Chlamydomonas reinhardtii (cc-2654, purchased from Chlamydomonas Genetics Center, Duck University, USA). TAP was selected as the medium for transgenic algae strains, and its preparation method was as follows: 25mL 4 × Beijerincksalts (4gMgSO 4 .7H2 O, 2gCaCl 2 .2H 2 O, 16gNH 4 Dissolve Cl in deionized water to 1L), 2.42g Tris, 1mL 1M(K)PO 4 , 1mLTrace trace element mixture (50gNa 2 EDTA, 1.1g (NH 4 ) 6 Mo 7 o 24 , 1.57gCuSO 4 .5H 2 O, 1.61gCoCl 2 .6H 2 O, 4.99gFeSO 4 .7H 2 O, 22gZnSO 4 .7H 2 O, 5.6gMnCl 2 .4H 2 O, 11.4gH 3 BO 3 Dilute to 1L deionized water), mix the above solutions and di...

Embodiment 3

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

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

[0079] From Example 1, it can be known that the plasmid pUC57HAV1 contains the HAV1 gene, and the pUC57HAV1' contains the HAV1' gene. Due to the addition of an appropriate restriction enzyme site while optimizing the sequence, it can be obtained by double restriction enzymes NheI and PmacI. HAV1 and HAV1' genes were obtained by enzyme digestion (purchased from Fermentas). 用EcoRⅠ消化质粒pH105(WuJXetal.Efficientexpressionofgreenfluorescentprotein(GFP)mediatedbyachimericpromoterinChlamydomonasreinhardtii.ChineseJournalofOceanologyandLimnology.Vol.26No.3,P.242-247,2008)得到含有HSP70A-RBCS2启动子和RBCS2终止子的部分约731bp,插入到pSP124( In Lambrerasv, StevensDR, PurtonS. Efficient foreign gene expression in Chlamydomonasreinhardtiimediatedbyanendogenousintron. Pl...

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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 have shown that for Asia alone, a relatively mild outbreak of bird flu caused a loss of US$90-110 billion. If it is a global bird flu outbreak, it may cause a global economic recession. At the same time, according to estimates by economists, when bird flu breaks out, the direct economic losses caused by bird flu to trade and tourism in the affected areas cannot be measured in numbers. Among 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 A2 (H2N2) virus that broke out in 1957. The spread of subtype influenza virus and the spread of A3 (H3N2) subtype influenza that broke out...

Claims

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Application Information

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Patent Type & Authority Patents(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
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