Construction method and use of grass carp matrix metalloproteinase inhibitor-2 gene, encoded recombinant protein and its spatiotemporal expression map

A protease inhibition and matrix metal technology, applied in the field of life sciences, can solve the problems of many grass carp diseases and losses in aquaculture

Inactive Publication Date: 2011-12-07
SHANGHAI OCEAN UNIV
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Problems solved by technology

However, grass carp suffers from many diseases, among which the hemorrhagic disease of grass carp is the worst, which brings huge losses to the aquaculture industry.

Method used

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  • Construction method and use of grass carp matrix metalloproteinase inhibitor-2 gene, encoded recombinant protein and its spatiotemporal expression map
  • Construction method and use of grass carp matrix metalloproteinase inhibitor-2 gene, encoded recombinant protein and its spatiotemporal expression map
  • Construction method and use of grass carp matrix metalloproteinase inhibitor-2 gene, encoded recombinant protein and its spatiotemporal expression map

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Embodiment Construction

[0024] The following specific embodiments of the present invention are only limited to the description of the present invention, and constitute limitations on the protection scope of the present invention.

[0025] 1. Sequence Alignment

[0026] According to a report by Iron in 1995, as long as the E-value value between the two sequences is less than 0.005 after comparison with BLAST software, there is absolute homology between the two sequences, which proves that the sequences are the same gene sequence. According to the EST sequence marked by the grass carp head and kidney cDNA library in our laboratory, BLAST search was carried out in NCBI. The partial coding region and 3' terminal nucleotide sequence of grass carp TIMP-2 gene were obtained. Gene-specific primers were designed based on this sequence.

[0027] 2. Grass carp TIMP-25' end clone

[0028] Grass carp total RNA extraction: Grass carp total RNA was extracted using the tissue RNA extraction reagent from TaKaRa Co...

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Abstract

The invention discloses the construction method and application of the grass carp matrix metalloproteinase inhibitory factor-2 gene, the encoded recombinant protein and its spatiotemporal expression map. The nucleotide sequence of the grass carp matrix metalloproteinase inhibitor-2 gene of the present invention is shown in SEQ ID NO:1. The amino acid sequence of the recombinant protein encoded by the nucleotide sequence is shown in SEQ ID NO:2. The matrix metalloproteinase inhibitor-2 gene of grass carp of the present invention is expressed in grass carp tissue and development process, and this gene is transcribed in a large amount in blood, mesonephros, head kidney, spleen, gill, intestine, heart, in brain, muscle, liver , There is a small amount of transcription in the skin, and the existence of a small amount of mRNA is detected in the fin rays. The gene was transcribed in a large amount from the caudal fin stage, and reached the maximum expression level on the 3rd day of fry. The invention also discloses the application of the grass carp matrix metalloproteinase inhibitor-2 gene and the coded recombinant protein in raising grass carp seedlings and as a feed additive for grass carp.

Description

technical field [0001] The invention belongs to the technical field of life sciences, and in particular relates to a grass carp matrix metalloproteinase inhibitor of metalloproteinases-2 (grass carp Tissue inhibitor of metalloproteinases-2, gcTIMP-2) gene, a recombinant protein encoded and a construction method for its temporal and spatial expression. Application of grass carp matrix metalloproteinase inhibitor-2 (grass carp Tissue inhibitor of metalloproteinases-2, gcTIMP-2) gene. Background technique [0002] Grass carp is a kind of herbivorous fish with low feeding cost and wide breeding area. It is the main species of freshwater aquaculture in my country, and its output accounts for about 20% of the total freshwater aquaculture output. However, there are many diseases of grass carp, among which hemorrhagic disease of grass carp is the worst, which brings huge losses to the aquaculture industry. At present, the cause of fulminant hemorrhagic disease in aquatic animals ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/15C07K14/81C12Q1/68A01K61/00A23K1/16A23K1/18A23K20/147A23K50/80
CPCY02A40/81
Inventor 李家乐徐晓雁沈玉帮宣云峰王荣泉
Owner SHANGHAI OCEAN UNIV
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