Molt-inhibiting hormone-1 protein of mitten crab

A technology of molting inhibitory hormone and Chinese mitten crab, applied in the field of genetic engineering, can solve the problems of small individuals, low market prices, economic losses of producers, etc.

Inactive Publication Date: 2006-01-25
NANJING NORMAL UNIVERSITY
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Problems solved by technology

Moulting is an important part of the growth of river crabs. The periodic molt of the exoskeleton is necessary for the growth of river crabs. However, during the breeding process, some biennial river crabs have failed to molt and become stiff crabs. The individuals are small and the market price is low. cause large economic losses

Method used

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  • Molt-inhibiting hormone-1 protein of mitten crab
  • Molt-inhibiting hormone-1 protein of mitten crab
  • Molt-inhibiting hormone-1 protein of mitten crab

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Embodiment Construction

[0108] (1) Preparation of molting inhibitory hormone 1 (Ers-MIH 1) fusion protein of Chinese mitten crab

[0109] 1. Using the extracted total RNA of eye stalk of Chinese mitten crab as a template, use the Superscript First-Strand Synthesis System for RT-PCR kit to reverse transcribe and synthesize the first-strand cDNA.

[0110] 2. Use the primers ErsM1 (5'-ccggattcATGGGAATCATCAACGCCGAG-3') and ErsM2 (5'-cgctcgagTTATTGCCCGAGGATGCT-3') to amplify the cDNA of the Ers-MIH 1 mature peptide using the first strand cDNA as a template, and the reaction conditions are: 94°C Pre-denaturation for 3 min, denaturation at 94°C for 30 s, annealing at 54°C for 30 s, extension at 72°C for 1 min, 30 cycles, and extension at 72°C for 10 min.

[0111] 3. After the above amplified product was purified by tapping, it was connected to the pMD18-T vector (Dalian Bao Biology) to construct the recombinant plasmid pMD18-T-MIH 1 and transform it into DH5α competent cells. The positive clones screened o...

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Abstract

The present invention belongs to the field of gene engineering technology. The Eriocheir sinensis molt-inhibiting hormone-1 (Ers-MIH 1) gene has length of 3506 bp, includes 3 exons, 2 introns, 5'-end upstream control region of 412 bp and 3'-end UTR of 917 bp. The polypeptide derived based on Ers-MIH 1 gene sequence consists of 75 amino acid mature peptides and 35 amino acid signal peptides. These mature peptides has Eriocheir sinensis molt-inhibiting function and has important control effect on the growth of Eriocheir sinensis. The Ers-MIH 1 gene has GenBank search number of AY310313. The present invention utilizes the Ers-MIH 1 gene in cloning fusion protein, the fusion protein is used in immunizing animal to prepare polyclonal antibody, and the antibody or the gene expressed control is used in solving the problem of abnormal molt causing small body of Eriocheir sinensis.

Description

1. Technical field [0001] The invention belongs to the technical field of genetic engineering. 2. Background technology [0002] Chinese mitten crab (Eriocheir japonica sinensis), also known as river crab, hairy crab, and hairy crab, is an important economic crab in my country. Moulting is an important part of the growth of river crabs. The periodic molt of the exoskeleton is necessary for the growth of river crabs. However, during the breeding process, some biennial river crabs have failed to molt and become stiff crabs. The individuals are small and the market price is low. cause greater economic losses. The main function of molt-inhibiting hormone (MIH) is to regulate the growth of crustaceans by inhibiting the synthesis of Y-organ ecdysone. 3. Contents of the invention [0003] The sequence of the Chinese mitten crab molting inhibitory hormone 1 (Ers-MIH 1) gene described by the inventor was obtained by the inventor through a series of experiments at the Institute of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C07K16/18A61K38/16A61K48/00A61P43/00
Inventor 周开亚宋霞马长艳郭豫杰
Owner NANJING NORMAL UNIVERSITY
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