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文蛤抗弧菌性状相关MITF基因及其多态性的鉴定

Identification of an MITF gene and its polymorphisms associated with the Vibrio resistance trait in the clam Meretrix petechialis.

作者信息

Zhang Shujing, Yue Xin, Jiang Fengjuan, Wang Hongxia, Liu Baozhong

机构信息

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; University of Chinese Academy of Sciences, Beijing 100049, China.

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.

出版信息

Fish Shellfish Immunol. 2017 Sep;68:466-473. doi: 10.1016/j.fsi.2017.07.035. Epub 2017 Jul 19.

Abstract

Microphthalmia-associated transcription factor (MITF) regulates the transcription of its target genes by binding to their promoters. In this study, an MITF gene, MpMITF was identified in the clam Meretrix petechialis. The full-length cDNA of MpMITF is 3564 bp with an ORF of 1365 bp. The deduced amino acid sequence consists of a conserved functional structure of bHLH-LZ, which could bind with E-box. The mRNA and protein expression levels of MpMITF were significantly up-regulated 6 h post-Vibrio injection. The mRNA expression of MpMITF increased on day 2 and peaked on day 10 post-Vibrio immersion. Furthermore, MpMITF expression was significantly up-regulated in most resistant families of clams (P < 0.05) but did not change significantly in most susceptive families of clams after the Vibrio immersion challenge. These results suggest that, in clams, MpMITF participates in the immune response against a Vibrio infection. Genotyping in two clam groups with different resistant levels to Vibrio parahaemolyticus (i.e., 11-R and 11-S), thirteen SNPs and five haplotypes were detected in the DNA sequence of MpMITF, of which five SNPs and two haplotypes were associated with Vibrio resistance. Four SNPs (SNP2, 5, 6 and 13) and one haplotype (Hap1) were further confirmed to be associated with Vibrio resistance in M. petechialis by association analysis in different clam families. This study deepens the understanding of MITF in marine bivalves and provides potential candidate markers for resistance selection in the clam M. petechialis.

摘要

小眼症相关转录因子(MITF)通过与靶基因的启动子结合来调控其转录。在本研究中,在文蛤(Meretrix petechialis)中鉴定出一个MITF基因,即MpMITF。MpMITF的全长cDNA为3564 bp,开放阅读框为1365 bp。推导的氨基酸序列包含一个保守的bHLH-LZ功能结构,该结构可与E-box结合。注射弧菌后6小时,MpMITF的mRNA和蛋白表达水平显著上调。弧菌浸泡后第2天,MpMITF的mRNA表达增加,并在第10天达到峰值。此外,在弧菌浸泡攻击后,MpMITF在大多数抗性较强的文蛤家系中表达显著上调(P < 0.05),但在大多数易感家系中变化不显著。这些结果表明,在文蛤中,MpMITF参与了针对弧菌感染的免疫反应。在对副溶血性弧菌具有不同抗性水平的两个文蛤群体(即11-R和11-S)中进行基因分型,在MpMITF的DNA序列中检测到13个单核苷酸多态性(SNP)和5个单倍型,其中5个SNP和2个单倍型与弧菌抗性相关。通过对不同文蛤家系的关联分析,进一步证实了4个SNP(SNP2、5、6和13)和1个单倍型(Hap1)与文蛤对弧菌的抗性相关。本研究加深了对海洋双壳贝类中MITF的理解,并为文蛤抗性选择提供了潜在的候选标记。

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