凡纳滨对虾(Litopenaeus vannamei)Wnt9α基因的克隆与表达分析

Cloning and Expression Profile Analysis of the Wnt9α Gene from Litopenaeus vannamei

Wnt信号通路已被证实在细胞增殖分化、胚胎发育等多方面起重要作用,Wnt基因家族是重要的调节因子。为了了解更多的无脊椎动物Wnt基因相关的信息,本研究首次克隆并鉴定了凡纳滨对虾（Litopenaeus vannamei）中的Wnt9α基因,命名为Lv Wnt9α,其基因全长c DNA为1 845 bp,包含长度为1 383 bp的开放阅读框（open reading frame,ORF）,编码460个氨基酸,包括一个信号肽和一个Wnt1结构域。实时荧光定量PCR（RT-PCR）分析表明Lv Wnt9α在被检测的所有组织中都有表达,其中在眼柄、肝胰腺、后盲囊、胃的表达量较高。白斑综合征病毒（white spot syndrome virus,WSSV）感染后,凡纳滨对虾血液中Lv Wnt9α基因的表达量呈先上升后下降趋势,在感染后48 h达到最高。革兰氏阴性菌副溶血弧菌（Vibrio parahaemolyticus）和革兰氏阳性菌金黄色葡萄球菌（Staphylococcus aureus）感染后,凡纳滨对虾血细胞中Lv Wnt9α基因表达量均在24 h达到最高水平,分别为对照组表达量的6.8倍和6.2倍。所有实验结果表明,Lv Wnt9α基因可能参与凡纳滨对虾先天性免疫应答途径。

Wnt gene family is the important regulatory factor of Wnt signaling,which has been proved that plays a vital role in cell proliferation,differentiation and embryonic development.To know more about the Wnt genes in inveterates,a Wnt gene in Litopenaeus vannamei named Lv Wnt9α was studied in this paper.The full-length c DNA of Lv Wnt9α was 1 845 bp with an open reading frame of 1 383 bp,which encoded a putative protein of 460 amino acids including a signal peptide and a Wnt1 domain.The results of Real-time PCR（RT-PCR） demonstrated that Lv Wnt9α was expressed in all the tested issues and was abundant in eyestalk,hepatopancreas,pyloric caecum and stomach.Upon WSSV infection,the expression levels of Lv Wnt9α in hemocyte increased at first and then declined,and it reached the peak at 48 h.After infected with gram-negative bacterium Vibrio parahaemolyticus and gram-positive bacterium Staphylococcus aureus,Lv Wnt9α expression in hemocyte showed the highest level both at 24 h,which was 6.8-fold and 6.2-fold of the control,respectively.These results implied that Lv Wnt9α may play a role in the pathway of innate immune response in L.vannamei.