草鱼MHCI区基因的表达特征

发布时间：2018-02-26 21:03

本文关键词： MHC I区基因半定量PCR 组织表达实时荧光定量PCR 草鱼　出处：《华中农业大学学报》2017年05期 　论文类型：期刊论文

【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96h,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12h和24h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24h达最大值;PSMB9表达量在脾脏先上升后下降,在12h达最大值,在肾脏中则先下降后上升,在24h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24h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]:In order to fully understand the MHC I gene and its expression characteristics of grass carp, using known MHC I gene sequence of zebrafish, partial gene sequence of MHC I region of grass carp was obtained by local Blast. RT-PCR technique was used. The expression levels of BRD2OKNSL2RXRBTP-FABGLMHCMHC IaHPSMB9 and TAP2 in the tissues of healthy grass carp were detected. The results showed that the expression of FABGL was the highest in the kidney. The expression of other genes was the highest in blood. The expression of MHC I immune-related genes in spleen and kidney of grass carp was detected by real-time fluorescence quantitative PCR. The results showed that the expression of BRD2 was detected in spleen and kidney of grass carp after infection with Aeromonas hydrophila. The expression of MHC Ia in kidney and spleen reached the highest level at 12 h and 24 h, respectively. The expression of MHC Ia in spleen and kidney increased first and then decreased. The expression of PSMB9 increased first in the spleen and then decreased in the spleen, reached the maximum at 12 h, then increased in the kidney, and reached the maximum at 24 h. The total expression of TAP2 decreased first and then increased in the spleen and kidney at 24 h, and reached the maximum at 24 h. In the end, the expression of .BRD2MHC IaMMB9 / TAP2 was significantly changed after the attack, indicating that these four genes play an important role in grass carp immunity.
【作者单位】：华中农业大学水产学院/农业部淡水生物繁育重点实验室/淡水水产健康养殖湖北省协同创新中心;
【基金】：中央高校基本科研业务费专项(2662015PY041;2015PY134)
【分类号】：S917.4

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