草鱼MHC I区基因的表达特征
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中央高校基本科研业务费专项(2662015PY041;2015PY134)


Expression patterns of MHC I region in grass carp, Ctenopharyngodon idella
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    摘要:

    为全面了解草鱼MHC I区基因及其表达特征,利用已知斑马鱼MHC I区基因序列,通过本地Blast得到草鱼MHC I区的部分基因序列。采用RT-PCR技术,检测MHC I区BRD2、KNSL2、RXRB、TAPBP、FABGL、MHC Ia、PSMB9以及TAP2等8个基因在健康草鱼各组织的表达水平。结果表明,除FABGL在肾脏表达量最高外,其他基因都在血液中的表达量最高。通过实时荧光定量PCR检测草鱼在嗜水气单孢菌感染后4、12、24、48及96 h,MHC I区免疫相关基因在脾脏和肾脏中的表达情况。结果发现,BRD2表达量分别在12 h和24 h在肾脏和脾脏中达到最高;MHC Ia表达量在脾脏和肾脏中都先上升后下降,并都在24 h达最大值;PSMB9表达量在脾脏先上升后下降,在12 h达最大值,在肾脏中则先下降后上升,在24 h达最大值;TAP2在脾脏和肾脏中整体呈先下降后上升的趋势,在24 h都达最大值,并最后都回调至正常范围。BRD2、MHC Ia、PSMB9、TAP2的表达量在攻毒后都有显著性变化,表明这4个基因在草鱼免疫方面具有重要的作用。

    Abstract:

    The partial cDNA sequences of MHC I genes were obtained from grass carp (Ctenopharyngodon idella) genome sequence based on local Blast against zebrafish's MHC I region. Gene expression levels of BRD2, KNSL2, RXRB, TAPBP, FABGL, MHC Ia, PSMB9 and TAP2 in various tissues were detected by semi-quantitative RT-PCR. The results showed that expression of 7 genes above were highest in the blood but that of FABGL was highest in the kidney. After Aeromonas hydrophila infection at 4, 12, 24, 48 and 96 h, expression level of immune-related MHC I genes was detected in the spleen and kidney with quantitative realtime PCR(qRT-PCR). The expression level of BRD2 gene reached the peak at 12 h in the kidney but at 24 h in the spleen. The expression level of MHC Ia gene increased first and then decreased both in the spleen and kidney, reaching the maximum at 24 h. In the spleen, the expression of PSMB9 increased signicantly and reached the peak at 12 h, and then decreased signicantly at 48 h, following by a recovery to normal level at 96 h; in the kidney, the expression peaked at 24 h, and then continued to decline. The expression levels of TAP2 gene were first down regulated and then up regulated in the spleen and kidney, and backed to normal level at last. It is indicated that MHC I genes are related to the immune of grass carp, laying a foundation for further studying fish diseases and breeding resistance cultivars.

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吴凡,郭稳杰,詹柒凤,张锋,胡晓坤,梁旭方,刘红.草鱼MHC I区基因的表达特征[J].华中农业大学学报,2017,36(5):60-66

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  • 收稿日期:2016-10-30
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  • 在线发布日期: 2017-07-20
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