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引用本文:任毅鹏,高 晶,潘宝平,高 虹,闫春财.青蛤(Cyclina sinensis)TLR2基因的克隆与表达分析.海洋与湖沼,2014,45(5):1037-1043.
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青蛤(Cyclina sinensis)TLR2基因的克隆与表达分析
任毅鹏,高晶,潘宝平,高虹,闫春财
天津师范大学生命科学学院,天津师范大学生命科学学院,天津师范大学生命科学学院,天津师范大学生命科学学院,天津师范大学生命科学学院
摘要:
利用转录组文库分析得到青蛤(Cyclina sinensis)的Toll样受体2(Toll-like receptor 2, TLR2)基因序列, 采用荧光定量PCR法分析该基因的表达过程。结果显示, 青蛤TLR2基因的cDNA开放阅读框为2082bp, 编码693个氨基酸, 具典型的LRRs、单次跨膜区和TIR结构域。该基因在血淋巴中的表达量最高, 与肝脏、鳃、外套膜、闭壳肌和性腺有显著性差异(P<0.05)。在鳗弧菌和Poly I:C刺激下, 青蛤血淋巴中TLR2基因3h开始升高, 48h达到最大值, 与对照组和空白组有极显著性差异(P<0.01); 藤黄微球菌刺激下, 血淋巴中CsTLR2基因3h开始升高, 48h亦达到最大值, 实验组与对照组有显著性差异(P<0.05)。
关键词:  青蛤  转录组文库  Toll样受体2  荧光定量PCR
DOI:10.11693/hyhz20140600183
分类号:
基金项目:天津市科委应用基础与前沿技术重点项目资助, 12JCZDJC22800 号, 09JCZDJC19300 号
附件
EXPRESSION AND CLONING OF TOLL-LIKE RECEPTOR 2 GENE FROM CYCLINA SINENSIS
Ren Yipeng,Gao Jing,Pan Baoping,Gao Hong and Yan Chuncai
College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University
Abstract:
The gene CsTLR2 (Cyclina sinensis toll-like receptor 2) were cloned and analyzed with transcriptome library. The expression of CsTLR2 was detected in real-time quantitative PCR. Results show that the open reading frame of CsTLR2 consisted of 2082bp, encoding 693 amino acids with the typical leucine-rich repeats, a single transmembrane area, and toll/interleukin-1 homologous region. Gene CsTLR2 was expressed in the highest level in hemolymph significantly different from those in liver, adductor muscle, gills, mantle, and gonad (P<0.05). The expression of TLR2 in C. sinensis hemolymph injected with Vibrio anguillarum and Poly I:C increased in 3h and reached the highest level in 48h, which is very significantly different (P<0.01) from those of the control group and the blank group. The expression of TLR2 in C. sinensis hemolymph injected with Micrococcus luteus increased in 3h and reached the highest level in 48h and it was significantly different (P<0.05) from that of control group.
Key words:  Cyclina sinensis  transcriptome library  Toll-like receptor 2  real-time PCR
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