生物技术通报 ›› 2015, Vol. 31 ›› Issue (3): 199-206.doi: 10.13560/j.cnki.biotech.bull.1985.2015.04.029

• 研究报告 • 上一篇    下一篇

松江鲈(Trachidermus fasciatus)凝血因子XI的基因克隆与表达分析

齐琪, 亓云月, 杨慧, 毕彩红, 韩晓弟,   

  1. (山东大学,威海 264209)
  • 收稿日期:2014-08-26 出版日期:2015-03-16 发布日期:2015-03-16
  • 作者简介:齐琪,研究方向:鱼类免疫;E-mail:530287931@qq.com
  • 基金资助:
    威海市科委项目(1070432121313)

Songjiang Sea Bass(Trachidermus fasciatus)Analysis of Gene Cloning and Expression of Coagulation Factor XI

Qi Qi Qi Yunyue Yang Hui Bi Caihong Han Xiaodi   

  1. (Shandong University, Weihai 264209)
  • Received:2014-08-26 Published:2015-03-16 Online:2015-03-16

摘要: 克隆松江鲈凝血因子XI基因cDNA序列,分析表达模式。利用RACE 技术从松江鲈中克隆获得了凝血因子XI的cDNA全长序列(命名为TfXI),并对其进行生物信息学和表达模式分析。获得TfXI cDNA全长1 287 bp,包括13 bp 的5'端非编码区,1 143 bp的开放阅读框以及131 bp 的3'端非编码区。开放阅读框编码280个氨基酸的多肽链,预测的蛋白大小为42.9 kD。N端含有由第22-105位氨基酸,112-196位氨基酸、205-279位氨基酸和289-369位氨基酸形成的4个串联排列的典型APPLE结构域。NCBI Blast结果显示TfXI与其他物种凝血因子XI的相似性为30%-63%,进化树分析显示,TfXI符合传统进化规律。Real time PCR分析表明,经LPS刺激96 h后,松江鲈脾脏TfXI表达量明显提高(P< 0.01)。

关键词: 松江鲈, 凝血因子XI, 基因克隆, Real time PCR

Abstract: It was to clone the full length cDNA encoding coagulation factor XI(FXI)of Trachidermus fasciatus. A TfXI gene from Trachidermus fasciatus(TfXI)was cloned and characterized by RACE technology. The TfXI cDNA composed of 1 287 bp with a 13 bp of 5'-UTR, 1 143 bp open reading frame(ORF)and 131 bp 3'-UTR, encoded a polypeptide of 280 amino acids. Sequence alignment of TfXI showed the highest similarity of 63% with Haplochromis burtoni FXI protein. After LPS stimulation, transcripts of TfXI were significantly increased and reached to peak at 96 h p.i. It indicated that TfXI may play an important role in immune response of T. fasciatus during pathogen challenge.

Key words: Trachidermus fasciatus, coagulation factor XI, cloning, real time PCR