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A kind of compound crucian carp itln protein and its preparation method and application

A protein and compound technology, applied in the field of genetic engineering research and development, can solve the problems of healthy fish breeding and water pollution, and achieve the effects of enriching the gene pool, high economic value and reducing production costs.

Inactive Publication Date: 2020-11-24
HUNAN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to rapid climate change and seriously polluted water quality problems, it has dealt a serious blow to fish healthy farming, and it is also a problem that is difficult to solve

Method used

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  • A kind of compound crucian carp itln protein and its preparation method and application
  • A kind of compound crucian carp itln protein and its preparation method and application
  • A kind of compound crucian carp itln protein and its preparation method and application

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Experimental program
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Embodiment

[0046] 1. Obtain the ORF of the ITLN gene of Hefang crucian carp by designing merger primers:

[0047] According to the conservation of ITLN amino acid sequence in various species, degenerate primers were designed. The first primer is ITLN-F1: 5'-ATGCTGCGCTTCATTTTCTTCCT (see SEQ ID NO: 3), the second primer is ITLN-R1: 5'-TTAACGGTAGAAGAGCAGCACGG (see SEQ ID NO: 4), reverse transcription with M-MLV Enzyme kit (Promega, Madison, WI, USA) reverse-transcribed the Hefang crucian carp liver cDNA as a template, and obtained the PCR amplification product through touchdown PCR (touchdown PCR) technology, which was sequenced and compared, and determined to be The ITLN gene of Hefang crucian carp is 945bp in length (see SEQ ID NO: 1), and its amino acid sequence is deduced (see SEQ ID NO: 2). The PCR reaction conditions were: pre-denaturation at 94°C for 4 min; denaturation at 94°C for 30 s, annealing at 72°C for 60 s, a total of 5 cycles; denaturation at 94°C for 30 s, annealing at 65°...

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Abstract

The invention discloses a WR ITLN protein. The protein is a protein with an amino acid sequence as shown in SEQ ID NO:2, or is a protein which has the same or higher activity and is obtained by substituting, deleting and / or adding one or more amino acids to an amino acid sequence as shown in SEQ ID NO:2 and / or carrying out terminal modification. The WR ITLN protein is discovered, not only enrichesa gene pool of WR, but also can be applied to preparation of recombinant protein, is further applied to the inhibition of growth of aeromonas hydrophila and / or streptococcus agalactiae and the preparation of immune preparations or feed additives for aquatic animals, and provides a new practical basis for physiological immune research of the WR. The invention further discloses a preparation methodof the WR ITLN protein, and application of the WR ITLN protein in the inhibition of growth of aeromonas hydrophila and / or streptococcus agalactiae and the preparation of immune preparations or feed additives for aquatic animals.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering research and development, and in particular relates to a combined crucian carp Intelectin (ITLN) protein and a preparation method thereof and its ability to inhibit the growth of Aeromonas hydrophila and / or Streptococcus agalactiae, and to prepare immune preparations for aquatic animals or application in feed additives. Background technique [0002] Intestinal lectin (Intelectin, ITLN), belonging to the lectin family, is a new type of soluble β-galactofuranose lectin. Mammalian ITLN was first discovered in Paneth cells of the small intestine of mice, and there are two isoforms of ITLN in mice and humans. Vertebrate ITLN mainly contains an N-terminal fibrinogen-like-domain (FReD) and a C-terminal intestinal lectin domain (intelectin domain). In general, the C-terminal β and γ chains of ITLNs are conserved, while the N-terminal has different modifications. The trimeric structure forme...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/46C12N15/70C12N15/66A23K50/80A23K20/147A61K38/17A61P31/04
CPCA23K20/147A23K50/80A61K38/00A61P31/04C07K14/461C12N15/66C12N15/70
Inventor 罗盛伟刘少军罗凯坤覃钦博张纯陶敏刘庆峰胡方舟王石
Owner HUNAN NORMAL UNIVERSITY
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