Chlamys farreri alpha2 macroglobulin gene and its preparing process

A technology of macroglobulin and Chlamys farreri, applied in botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve problems such as economic losses, threats to sustainable development of industries, and death of Chlamys farreri

Inactive Publication Date: 2004-11-03
OCEAN UNIV OF CHINA
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Problems solved by technology

However, with the aggravation of breeding diseases in recent years, large-scale death of scallops during the grow-out period

Method used

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  • Chlamys farreri alpha2 macroglobulin gene and its preparing process
  • Chlamys farreri alpha2 macroglobulin gene and its preparing process
  • Chlamys farreri alpha2 macroglobulin gene and its preparing process

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Embodiment Construction

[0009] According to the α2M gene sequence of the arthropod horseshoe crab (Limulus polyphemus), the guessing primers were selected in the α2M receptor binding region: α2MUP (5'-GGT TGT GGT GAA CAA AACATG G-3') and α2MDN (5'-GTA TCC ATC CAG AAA CCA TCT T-3'). Total RNA of blood cells of Chlamys farreri was prepared and reverse transcribed into cDNA. Take 2 μL of the above cDNA as a template for polymerase chain reaction (PCR), the conditions are: 94°C, 2min, 1 cycle; 94°C 30sec, 57.5°C 30sec, 72°C 1min, 30 cycles; 72°C 10min, 1 cycle. A 1172bp fragment of the corresponding receptor region was obtained. Design the degenerate primer BAIT (5'-AAG CCC GTTGTA GAA / G ATI A / C G-3') in the α2M decoy region and the nested primer OUTR (5'-TTC TGTCTC AAT CTT GCT GG-3 ') and INR (5'-TGT TGG TGA GAT ACT GAAGG-3') combined for nested reverse transcription polymerase chain reaction (RT-PCR), the conditions are: 94°C 1.5min, 1 cycle; 94°C 30sec , 50°C 45sec, 72°C 1min, 35 cycles; 72°C 10min,...

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Abstract

The alpha-2 macroglobulin gene of Chlamys Farreri features that it encodes macroglobulin polypeptide of Chlamys Farreri with SEQ ID No. 2 sequence. First, guessmer primer and degenerate primer are made to produce inverse transcriptive PCR, and then, fast cloning in 3' end and 5' end is performed. The alpha-2 macroglobulin gene of Chlamys Farreri is used in culture and development of excellent resistant Chlamys Farreri varieties.

Description

technical field [0001] The invention relates to an α2 macroglobulin gene of Chlamys farreri and a preparation method thereof. Background technique [0002] α2-macroglobulin (α2M, abbreviated as α2M) is a macromolecular glycoprotein in animal plasma. The activity of derived proteases is widely involved in a variety of physiological functions. When inflammation occurs, the body can induce the synthesis of abundant α2 macroglobulin, which enters the tissue, combines with the excess protease released by the tissue, and removes the latter, preventing further damage to the tissue. α2M can also reduce the damage to the host and enhance the host's disease resistance by inhibiting various proteases and toxins released by parasites and pathogens that invade the body. It is an important non-specific immune factor. Chlamys farreri belongs to the phylum Molluscs, Phallobranches, Pearl Oysters, and Scallopidae. It is mainly distributed in the coasts of China, North Korea, South Korea, a...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12P19/34
Inventor 马洪明麦康森徐玮刘付志国张文兵谭北平
Owner OCEAN UNIV OF CHINA
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