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Complement C3 gene, carrier, recombination strain and protein of Epinephelus coioids and application thereof

A grouper and protein technology, applied in the field of genetic engineering, can solve problems such as high morbidity and mortality, achieve high economic value, broad application prospects, and enrich the effect of gene pools

Active Publication Date: 2013-10-30
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the high morbidity and mortality caused by the current climate disasters and environmental pollution have always been difficult problems in the artificial breeding of grouper
There is no feed additive that can effectively enhance the immunity of grouper on the market

Method used

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  • Complement C3 gene, carrier, recombination strain and protein of Epinephelus coioids and application thereof
  • Complement C3 gene, carrier, recombination strain and protein of Epinephelus coioids and application thereof
  • Complement C3 gene, carrier, recombination strain and protein of Epinephelus coioids and application thereof

Examples

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Embodiment Construction

[0039] Obtaining the Middle Fragment of the Complement C3 Gene of the Grouper Plaques by Suppression Subtractive Hybridization

[0040] The low-temperature library of grouper was constructed by suppression subtractive hybridization (SSH), and some functional genes were sequenced, including fragments highly homologous to complement C3 of grouper. The length of the middle fragment of the complement C3 gene is 396bp (SEQ ID NO.5).

[0041] Synthesis of 5' fragment of complement C3 gene in grouper

[0042]The operation was performed according to the BD SMART RACE cDNA Amplification Kit (BD Bioscience Clontech, CA, USA) Protocol. According to the requirements of the kit, two downstream primers C351 and C352 were designed based on the sequence of the sequenced complement C3 middle fragment for nested PCR 。 The first downstream primer C351 is [5'-CTTTCGGCCTTGGCGGGTTTG] (SEQ ID NO.6), and the second downstream primer C352 is [5'-ACGGAAACAACACGTCCTCTACG] (SEQ ID NO.7).

[0043] ...

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Abstract

The invention discloses a complement C3 gene, a carrier, a recombination strain and protein of Epinephelus coioids and application thereof, wherein the amino acid sequence of the rC3B protein of garrupas is represented by SEQIDNO:4; or, the amino acid sequence represented by SEQIDNO:4 is substituted, deficient and / or increased by one or more amino acid and / or modified at the tail end; furthermore, the amino acid sequence has protein with same or higher activity. According to the invention, a complement C3 gene sequence of the garrupas and a functional fragment, namely an rC3B nucleotide sequence on the C3 sequence, are obtained for the first time; the gene bank of the garrupas is enriched; furthermore, the complement C3 gene can be applied to preparing recombination protein, fish immune preparation or feed additives; and a novel theory and practice foundation are provided for physiological immune research of the garrupas.

Description

technical field [0001] The present invention belongs to the technical field of genetic engineering, and specifically relates to a rC3B protein, a gene encoding the protein, a carrier containing the gene and applications thereof; and also relates to a complement C3 protein, a gene encoding the protein, a carrier containing the gene, and an application thereof. its application. Background technique [0002] The third complement component (C3) is the core of the complement system, which was discovered by Ritg in 1912 when he treated serum with snake venom. Among the components of complement, the serum content of complement C3 is the highest. Complement C3 protein has a special structure. It is a β2 glycoprotein composed of two peptide chains α and β. The two peptide chains are connected by disulfide bonds. The molecular weight is 195KDa, of which the α chain is 115KDa and consists of 998 amino acid residues The N-terminal and C-terminal are both serine; the β-chain is 75KDa, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/46C12N15/12C12N15/63A23K1/16
Inventor 王维娜祁增华刘玉凤
Owner SOUTH CHINA NORMAL UNIVERSITY
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