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Crassostrea hongkongensis antimicrobial peptide sa-lectin as well as recombinant expression method and application thereof

A technology of Hong Kong oyster and expression method, applied in application, antibacterial drugs, recombinant DNA technology, etc., can solve problems such as insufficient immunity of shellfish

Inactive Publication Date: 2016-08-17
QINGYUAN POLYTECHNIC
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  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to solve the existing problems such as insufficient immunity of shellfish at present. A sialic acid-binding lectin salectin gene was identified from the cDNA of Hong Kong oyster, and the full-length ORF of the modified gene was obtained by cloning

Method used

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  • Crassostrea hongkongensis antimicrobial peptide sa-lectin as well as recombinant expression method and application thereof
  • Crassostrea hongkongensis antimicrobial peptide sa-lectin as well as recombinant expression method and application thereof
  • Crassostrea hongkongensis antimicrobial peptide sa-lectin as well as recombinant expression method and application thereof

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Experimental program
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Embodiment 1

[0019] 1. RNA extraction

[0020] Total RNA was extracted using Trizol (Invitorgen), and the specific steps were as follows: (1) Take 50 mg of tissue in a mortar pre-cooled with liquid nitrogen, grind the tissue into powder, transfer it to a centrifuge tube containing 1 ml Trizol, and pipette , mix well; (2) Add 200 μL chloroform, shake vigorously for 40 seconds, and place at room temperature for 5 minutes; (3) Centrifuge at 12,000×g for 10 minutes at 4°C, transfer the supernatant to a new tube; (4) Add 0.5ml of isopropyl (5) Centrifuge at 4°C, 12,000×g for 10 min, and discard the supernatant; (6) Wash twice with 75% ethanol; (7) Discard the supernatant and add 100 μL DEPC Water dissolves RNA.

[0021] 2. cDNA library construction

[0022] The full-length cDNA library was constructed using the SMART cDNA library construction Kit (Clontech), and the specific steps were as follows: (1) Use MMLV to synthesize the first strand of cDNA; (2) Use LD PCR to amplify the cDNA; (3) Pro...

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Abstract

The invention discloses crassostrea hongkongensis antimicrobial peptide sa-lectin as well as a recombinant expression method and application thereof. The gene coding sequence of antimicrobial peptide sa-lectin is shown in SEQ ID NO: 1, in which 156 amino acids are coded, and is shown in SEQ ID NO: 2, in which 16 amino acids at the end N are signal peptide sequences. According to the crassostrea hongkongensis antimicrobial peptide sa-lectin, a prokaryotic expression vector for removing the signal peptide sequence is constructed, so that sa-lectin recombined proteins are expressed and purified and can suppress growth of various types of gram-positive bacteria and gram-negative bacteria; furthermore, by the combination of sialic acid, an important effect is achieved in immune defense of shellfish.

Description

technical field [0001] The invention relates to the technical field of shellfish medicines, in particular to a Hong Kong oyster antibacterial peptide sa-lectin and its recombinant expression method and application. technical background [0002] Shellfish farming occupies a pivotal position in the economic and social development of my country's coastal areas. It is one of the most important economic groups in the world and has great potential for development. Hong Kong Oysters ( Crassostrea hongkongensis ), belonging to the phylum Mollusca, Bivalvia, Pearl Oyster, and Oysteridae, is the most important shellfish economic animal in the coastal areas of South my country. However, the lack of good species leads to low immunity of oysters, disease outbreaks, and even mass mortality, which is the main bottleneck restricting the development of the shellfish industry. [0003] Therefore, how to improve the antibacterial ability of shellfish is an effective means to solve the curr...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/70A61K38/17A61P31/04A61P37/04C12R1/19
CPCC07K14/43509A61K38/00C12N15/70C12N2800/101
Inventor 冼昶华陈金辉
Owner QINGYUAN POLYTECHNIC
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