Larimichthys crocea scavenger receptor gene MARCO gene and immune defending function thereof

A technology of scavenger receptors and immune defense, which is applied in the field of genetic engineering, can solve problems such as incorrect recognition and binding of ligands, and achieve the effects of good total RNA extraction, integrity maintenance, and improved yield and purity

Inactive Publication Date: 2019-03-19
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that after knocking out the SRCR domain of human and mouse MARCO genes, they will not be able to correctly recognize and bind the corresponding ligands.

Method used

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  • Larimichthys crocea scavenger receptor gene MARCO gene and immune defending function thereof
  • Larimichthys crocea scavenger receptor gene MARCO gene and immune defending function thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024]大黄鱼清道夫受体基因MARCO基因,MARCO基因开放阅读框序列采用同源克隆法与RACE技术克隆获得,MARCO基因开放阅读框序列为如SEQ ID NO:1所示,具体为:ATGGAGACGTCGGTGGACCGCACCAACAACCGAGTCTCGTACACGCAGAGTAACCCGCTGTTCGACATGTCGCTCAGCAGGACTGACCTGTACAACTTCCAGCCTGACGATCTGAAGCCGGCGAGGCCGAGACGACAGTGGTGTTTAAACGTGATCATCGTTTACCTCGTCCTGCAGACCGGCCTGAACGCCTTCCTCATTTATAAAGTTTTCACGTTGGACTCATCGTCGTCGACCCTCGGGGCTCAGAAGCAGACGTCCAATCACATGCCTCAGGGAGGAGAACCGAGCGGCGATGACACCCTCCAAACTCTCGTCCGCAACAACAGTGAAGAAACCAAAACCCTGAGGGGTCAGCTGTGGGCGCTGGACAGCCAGGTCAAGAACCTCTGTGCAGAGGAGGGTCAGCTGGGCAGGATGAGGTCTGACCTGAAGATGCTCAACTCAAGCGCCCGAAACCTGGAGGACAAACTGATCAACCTCAAACAAGGACCTCCAGGTCCTCCGGGAAGAGACGGGCTTTCAGGACAGCCGGGCACACCTGGAGACAGAGGCCTCAAAGGTGACATCGGTGACGCTGGCCCTCAAGGACTAAAGGGTGACAACGGACAACCCGGACAGTCAGGAGCTCCGGGTCCCAGAGGACCACCTGGACCTGCAGGAGCCCCAGGGAATCAAGGACCAGGAGCTAAGGGAGAGAAAGGAGCACCTGGGGGCCCAGGCCTTCCTGGACCCAAAGGTGACCCCGGCACCGCCGGGCAAAAAGGATCTTCTGGATCTCAAGGTACCACTGGGCTACAAGGACAGAAAGCAGATTCTGGTCCACCGGGACCTCAAGGTGCATTGGGACCTCCTGGTGCCAAAGGAGATCAAGGTCCAGCTGGACCACCAGGTGCCAAAGCAG...

Embodiment 2

[0073] The large yellow croaker scavenger receptor gene MARCO gene, the MARCO gene is cloned by homologous cloning method and RACE technology, and the full-length cDNA sequence of the MARCO gene is 8275bp.

[0074] MARCO gene cloning includes the following steps:

[0075] 1) Total RNA extraction and detection;

[0076] 2) Synthesis of the first strand of cDNA;

[0077] 3) MARCO gene amplification and purification;

[0078] 4) T-A cloning.

[0079] Total RNA was extracted using Trizol Regent, guanidine phosphate and thiourea. Trizol Regent contained 0.035% guanidine phosphate and 0.015% thiourea.

[0080] Primers for gene amplification are:

[0081] MARCO–F: 5'-ATGGAGACGTAGGTGGAGGGCACC-3';

[0082] MARCO–R: 5'-CTAGGAGCCCTCCAGTGCCGG-3'.

[0083] The amplification reaction system is: 2.5 μL 10×PCR Buffer, 0.4 μL dNTPs, 0.8 μL MARCO-F, 0.8 μL MARCO-R, 0.6 μL Template cDNA, 0.4 μL Taq DNA polymerase (TaKaRa), 14.5 μL RNasefree dd H 2 O, the amplification conditions were: pre...

Embodiment 3

[0086] The large yellow croaker scavenger receptor gene MARCO gene, the MARCO gene is cloned by homologous cloning method and RACE technology, and the full-length cDNA sequence of the MARCO gene is 8275bp.

[0087] MARCO gene cloning includes the following steps:

[0088] 1) Total RNA extraction and detection;

[0089] 2) Synthesis of the first strand of cDNA;

[0090] 3) MARCO gene amplification and purification;

[0091] 4) T-A cloning.

[0092] Total RNA was extracted using Trizol Regent, guanidine phosphate and thiourea. Trizol Regent contained 0.04% guanidine phosphate and 0.02% thiourea.

[0093] Primers for gene amplification are:

[0094] MARCO–F: 5'-ATGGAGACGTAGGTGGAGGGCACC-3';

[0095] MARCO–R: 5'-CTAGGAGCCCTCCAGTGCCGG-3'.

[0096] The amplification reaction system is: 2.5 μL 10×PCR Buffer, 0.4 μL dNTPs, 0.8 μL MARCO-F, 0.8 μL MARCO-R, 0.6 μL Template cDNA, 0.4 μL Taq DNA polymerase (TaKaRa), 14.5 μL RNasefree dd H 2 O, the amplification conditions were: pre-d...

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Abstract

The invention discloses a larimichthys crocea scavenger receptor gene MARCO gene and an immune defending function thereof. An open reading frame sequence of the MARCO gene is obtained by adoption of ahomology-based cloning method and a RACE technique, the sequence is shown as SEQ ID NO:1, the overall length is 1218bp, and 405 amino acids are encoded. The MARCO gene is applied to the larimichthyscrocea immune defending function including congential immunity regulation for maintaining body health and evolution. The MARCO gene is obtained by adoption of the homology-based cloning method and theRACE technique, effectiveness in total RNA extraction in a cloning process is achieved, intrinsic RNA enzyme inactivation can be realized, and total RNA yield and purity are improved. By the immune defending function of the MARCO gene, a theoretical basis is provided for prevention of diseases caused by pathogenic bacteria, and theoretical guidance is provided for disclosing healthy aquaculture of larimichthys crocea.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the large yellow croaker scavenger receptor gene MARCO gene and its immune defense function. technical background [0002] large yellow croaker ( Larimichthys crocea ) is widely distributed in the southeast coast of my country, commonly known as "yellow croaker". The meat is tender and smooth, and has high nutritional value. After the market, the economic status of large yellow croaker has been continuously improved, and its artificial breeding has attracted more and more attention. However, the breeding of large yellow croaker is usually inseparable from the problem of resistance to pathogenic bacteria, so the development and utilization of immune-related genes is particularly important. Based on the completion of the whole genome sequencing of large yellow croaker, people's genetic research on large yellow croaker has reached a new level, and the characteristics of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/705C12N15/10
CPCC07K14/705C12N15/1096C12Q2531/113
Inventor 刘慧慧吴炯迟长凤黄伟
Owner ZHEJIANG OCEAN UNIV
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