Cloning of natural resistance associated macrophage protein2 (Sc-Nramp2)gene from sinonovacula constricta and the expression analysis under vibrio parahaemolyticus stimulation

Lin Dehai; Liu Chenshan; Dong Yinghui; He Lin; Lin Zhihua

doi:10.3969/j.issn.0253-4193.2017.08.007

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Lin Dehai, Liu Chenshan, Dong Yinghui, He Lin, Lin Zhihua. Cloning of natural resistance associated macrophage protein2 (Sc-Nramp2)gene from sinonovacula constricta and the expression analysis under vibrio parahaemolyticus stimulation[J]. Haiyang Xuebao, 2017, 39(8): 70-77. doi: 10.3969/j.issn.0253-4193.2017.08.007

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Lin Dehai, Liu Chenshan, Dong Yinghui, He Lin, Lin Zhihua. Cloning of natural resistance associated macrophage protein2 (Sc-Nramp2)gene from sinonovacula constricta and the expression analysis under vibrio parahaemolyticus stimulation[J]. Haiyang Xuebao, 2017, 39(8): 70-77. doi: 10.3969/j.issn.0253-4193.2017.08.007

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Cloning of natural resistance associated macrophage protein2 (Sc-Nramp2)gene from sinonovacula constricta and the expression analysis under vibrio parahaemolyticus stimulation

doi: 10.3969/j.issn.0253-4193.2017.08.007

Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China

Received Date: 2016-11-23
Rev Recd Date: 2017-04-13

Abstract

Abstract

The cDNA sequence of natural resistance associated macrophage protein in Sinonovacula constricta (Sc-Nramp2) was cloned by SMART RACE techniques. The ORF of cDNA sequence, coding for a protein of amino acids, was 1 773 bp and the molecular weight was deduced for 65.86 kDa. Sc-Nramp2 protein has the typical structural features with Solute carrier family 11 member, with 10 typical transmembrane domains and 2 glycosylation site. In 3'-UTR, Sc-Nramp2 was the presence of two IRE which was similar to the vertebrate Nramp2. Homology analysis indicated that the identity of Nramp with Crassostrea gigas was 71.6%. Real-time quantitative RT-PCR analysis indicated that the mRNA was expressed in all the six tissues, including adductor muscle, mantle, hepatopancreas, foot, waterpipe and gill. Nramp was expressed with the highest expression level in the hepatopancreas, then the gill, which had extremely significant differences from other tissues(P<0.01). The expression of Nramp in the hepatopancreas injected with V. Parahaemolyticus increased significantly(P<0.01). It rose to maximum expression at the 12^th hour and returned to the initial level. These results indicated that Sc-Nramp2 is involved in innate immunity of this species.
- Sinonovacula constricta,
- Sc-Nramp2,
- gene cloning,
- Vibrio Parahaemolyticus,
- gene expression

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References(35)

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