【摘要】 【目的】分析WDS基因在凡纳滨对虾(Litopenaeus vannamei)各组织中的表达情况及应对不同病原感染时的表达变化,以揭示WDS在动物体除生殖外其他生理活动中的功能作用。【方法】采用RACE-PCR克隆凡纳滨对虾WDS基因(LvWDS)cDNA序列,利用在线生物信息学软件进行序列分析,并以实时荧光定量PCR分析LvWDS基因在凡纳滨对虾不同组织中的表达情况及不同病原感染后的表达变化。【结果】LvWDS基因(GenBank登录号MH330316)c DNA序列全长1880 bp,其中,开放阅读框(ORF)978 bp,5’端非编码区(5’UTR)183 bp,3’端非编码区(3’UTR)719 bp,编码325个氨基酸。LvWDS氨基酸序列含有7个WD40重复序列(保守结构域),与拟穴青蟹(Scylla paramamosain)WDS氨基酸序列的相似性最高,为99%,基于WDS氨基酸序列相似性构建的系统发育进化树也显示LvWDS与拟穴青蟹WDS的亲缘关系最近。LvWDS基因在凡纳滨对虾精囊中的相对表达量最高,在血细胞、鳃和肠道等免疫组织中处于较高表达水平,在肌肉组织中的表达水平最低。经白斑综合症病毒(WSSV)和副溶血弧菌(Vibrio parahaemolyticus)感染后,凡纳滨对虾血细胞、鳃和肠道组织中LvWDS基因的相对表达量均不同程度发生变化。【结论】WDS进化十分保守,其在不同物种中的功能稳定。LvWDS除了在凡纳滨对虾生殖发育中发挥重要作用外,可能还与免疫相关,在凡纳滨对虾抗病免疫中发挥作用。更多还原

【Abstract】 【Objective】The purpose was to analyze the expression of WDS gene in tissues of Litopenaeus vannamei and variation in response to different pathogen infections,and reveal the function of will die slowly(WDS)in physiological activities except reproduction in animals.【Method】c DNA sequence of WDS gene(LvWDS)from L. vannamei was cloned by RACE-PCR. Bioinformatics analysis on the sequence was conducted,and distribution of LvWDS gene in various tissues as well as the expression variation after infection by different pathogens were studied by real-time fluorescence quantitative PCR.【Result】Full-length cDNA sequence of WDS gene from L. vannamei(Genbank accession number:MH330316)was 1880 bp with a 978 bp open reading frame(ORF)that encoded 325 amino acids. The 5’ non-coding region(5’UTR)was 183 bp while the 3’UTR was 719 bp. LvWDS amino acid sequence contained seven WD40 repeats(conserved domains)and shared the highest(99%)amino acid similarity with WDS amino acid sequence from Scylla paramamosain. The phylogenetic tree based on WDS amino acid sequence similarity indicated that LvWDS had the closest relationship with WDS in S. paramamosain.The expression of LvWDS gene was the highest in seminal vesicle and the lowest in mu-scle. The expression levels in immunity tissues such as hemocyte,gill and intestine were high. Upon white spot syndrome virus(WSSV)and Vibrio parahaemolyticus infection,the expression of LvWDS in hemocyte,gill,and intestine of L. vannamei was changed at varying degrees.【Conclusion】WDS evolution is very conservative,and maintains stable functions in different species. LvWDS plays an important role in reproduction of L. vannamei,and may relate to immunity and also functions in disease resistance and immunity of L. vannamei.更多还原