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FenneropenaeusChinensis Crustin gene and coded protein and cloning method therefor

A cloning method and gene coding technology, which is applied in the field of Chinese prawn Crustin gene and encoded protein and its cloning, can solve the problems of shrimp output value loss and achieve far-reaching effects of improving disease resistance

Inactive Publication Date: 2005-08-17
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the frequent outbreaks of prawn diseases in recent years, it has caused great losses to the output value of prawns.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1. A cloned Penaeus prawn Crustin gene has the following sequence:

[0021] Homologous gene one:

[0022] (1) Information of SEQ ID No 1 (see sequence listing)

[0023] (a) Sequence features:

[0024] * Length: 487 base pairs

[0025] * Type: nucleic acid

[0026] * Chain type: double chain

[0027] * Topology: Linear

[0028] (b) Molecular type: cDNA

[0029] (c) Assumption: No

[0030] (d) Antisense: no

[0031] (e) Original source: Chinese prawns (Fenneropenaeus chinensis)

[0032] (2) Information of SEQ ID No.2 (see sequence listing)

[0033] (a) Sequential features

[0034] * Length: 130 amino acids

[0035] * Type: amino acid

[0036] * Chain type: single chain

[0037] * Topology: Linear

[0038] (b) Molecule type: peptide

[0039] Sequence description: SEQ ID No.1

[0040] 1 A CTG GAG GCG ACC ATG AAG GGT CTC GGA GTC ATT CTG TTC TGC GTC 46

[0041] Met Lys Gly Leu Gly Val Ile Leu Phe Cys Val 11

[0042] 47 C...

Embodiment 2

[0064] Embodiment 2. The cloning method of Chinese prawn Crustin gene

[0065] 1. Total RNA extraction: Hemolymph was collected from the abdominal sinus at the base of the first abdominal segment of Penaeus chinensis with a syringe, and an equal volume of anticoagulant (27mM sodium citrate, 336mM NaCl, 115mM glucose, 9mM EDTA, pH7; Rodriguez et al. , 1995). Hemolymph was centrifuged at 4°C, 800g for 10min, and blood cells were collected for RNA extraction; total RNA was extracted using Unizol RNA extraction reagent from Shanghai Boxing Biological Company, and the extraction method was referred to the instruction manual.

[0066] 2. cDNA first-strand synthesis: according to the cDNA synthesis kit instructions of Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0067] 3. Shrimp Crustin cDNA fragment cloning

[0068] (1) Design primers according to the EST sequence obtained by large-scale sequencing of the cDNA library for PCR amplification to obtain the prawn Crus...

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PUM

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Abstract

The present invention relates to Crustin, and is especially one kind of Chinese prawn Crustin gene and its cloning process. The gene has the base sequence shown in SEQ ID No. 1 of the sequence list and the amino acid sequence shown in SEQ ID No. 2 of the sequence list. The cloning process includes: 1) extracting total RNA from the blood cell of Chinese prawn; 2) performing the first chain synthesis of cDNA; and 3) cloning with 3'-RACE and 5'-RACE to obtain Chinese prawn Crustin gene. The present invention clones one kind of Crustin gene from the blood cell of Chinese prawn for the first time, reveals all its coding sequences and non-coding sequences as well as its coded amino acid sequence, and makes it possible to prevent some prawn diseases caused by some pathogenic microbes.

Description

technical field [0001] The invention relates to Crustin, in particular to a Chinese prawn (Fenneropenaeuschinensis) Crustin gene and coded protein and a cloning method thereof. Background technique [0002] Shrimp farming is an important global economic industry and plays an important role in the marine aquaculture industry. However, due to frequent outbreaks of prawn diseases in recent years, great losses have been caused to the output value of prawns. The use of antibiotics has greatly curbed the prevalence of shrimp diseases and increased shrimp production. However, the ensuing emergence of drug-resistant bacteria, the destruction of the environment, and the harm to human health have attracted increasing attention, so the search for new antibacterial drugs has become increasingly urgent. [0003] Like other crustaceans, prawns do not have a specific immune system, but they have several highly effective non-specific immune methods to resist the invasion of foreign pathog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/10C12N15/29
Inventor 相建海柳峰松李富花董波
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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