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文蛤MIF基因多态性及其与抗弧菌能力的关联

The polymorphisms of a MIF gene and their association with Vibrio resistance in the clam Meretrix meretrix.

作者信息

Zou Linhu, 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 100039, China.

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.

出版信息

Dev Comp Immunol. 2016 Sep;62:116-26. doi: 10.1016/j.dci.2016.04.013. Epub 2016 Apr 19.

Abstract

Macrophage migration inhibitory factor (MIF) is an important proinflammatory cytokine that mediates both innate and adaptive immune responses. In this study, a homolog of MIF was identified in the clam Meretrix meretrix. Ten SNPs in the DNA partial sequence of MmMIF were found to be significantly associated with Vibrio resistance (P < 0.05). Distinct expression patterns of MmMIF among different haplotypes were observed after Vibrio challenge. The results showed that haplotypes did not affect MmMIF expression in the negative control group, while the expression of MmMIF in clams with Hap1 and Hap1/Hap2 was significantly lower than that with Hap2 at 24 h in the PBS-injected group but significantly higher than that with Hap2 in the Vibrio-injected group. The results also indicate that Hap1 and Hap1/Hap2 can specifically respond to mechanical stimulation while Hap2 can specifically respond to Vibrio infection. The effect of a missense mutation was detected by site-directed mutagenesis using fusion expression of the protein, which showed that the SNP g.737 (I > V) has no effect on redox activity and tautomerase activity. These studies identified a potential marker that is enriched in Vibrio-resistant clams that can be used for the genetic breeding of Meretrix meretrix.

摘要

巨噬细胞移动抑制因子(MIF)是一种重要的促炎细胞因子,介导先天性和适应性免疫反应。在本研究中,在文蛤中鉴定出了MIF的一个同源物。发现文蛤MIF(MmMIF)DNA部分序列中的10个单核苷酸多态性(SNP)与弧菌抗性显著相关(P < 0.05)。在弧菌攻击后,观察到不同单倍型间MmMIF有明显的表达模式。结果表明,在阴性对照组中,单倍型不影响MmMIF的表达,而在注射PBS组中,24小时时,具有单倍型1(Hap1)和单倍型1/单倍型2(Hap1/Hap2)的文蛤中MmMIF的表达显著低于具有单倍型2(Hap2)的文蛤,但在注射弧菌组中显著高于具有Hap2的文蛤。结果还表明,Hap1和Hap1/Hap2可特异性响应机械刺激,而Hap2可特异性响应弧菌感染。通过蛋白质融合表达的定点诱变检测到一个错义突变的影响,结果表明SNP g.737(I > V)对氧化还原活性和互变异构酶活性没有影响。这些研究鉴定出了一种在抗弧菌文蛤中富集的潜在标记,可用于文蛤的遗传育种。

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