Identification of immune-related genes from kidney and spleen of turbot, Psetta maxima (L.), by suppression subtractive hybridization following challenge with Vibrio harveyi.
Identification of immune-related genes from kidney and spleen of turbot, Psetta maxima (L.), by suppression subtractive hybridization following challenge with Vibrio harveyi.
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DOI：
10.1111/j.1365-2761.2008.00914.x
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发表时间：
2008-07
影响因子：
2.5
通讯作者：
Chunzai Wang;Xiao-Hua Zhang;A. Jia;Jixiang Chen;Brian Austin
中科院分区：
文献类型：
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作者：
Chunzai Wang;Xiao-Hua Zhang;A. Jia;Jixiang Chen;Brian Austin
利用抑制性消减杂交技术研究了大菱鲆Psetta maxima（L.）利用人工感染大菱鲆肾脏和脾脏mRNA构建的cDNA文库，对哈氏弧菌进行了研究。获得了49个表达序列标签。几个免疫系统基因被确定，包括主要组织相容性复合体（MHC）Ia类基因和热休克蛋白70基因。一些信号分子也存在于cDNA文库中，包括src家族酪氨酸激酶SCK，sgk-1丝氨酸-苏氨酸蛋白激酶和淀粉样蛋白受体样蛋白2。利用PCR技术从大菱鲆cDNA中克隆了MHC-Ia类基因的全长cDNA。大菱鲆MHC Class Ia的核苷酸序列已提交到GenBank，登录号为EF 032639。大菱鲆MHC Class Ia cDNA开放阅读框编码354个氨基酸，推导的氨基酸序列与比目鱼、青鳉、虹鳟、大西洋鳕、河豚、鸡、小鼠和人的同源性分别为68%、54%、51%、52%、57%、33%、29%和29%。对MHC Ia类基因进行定量逆转录酶-PCR，结果显示，在哈维氏弧菌攻击的大菱鲆中，MHC Ia类基因的表达水平增加到对照的四倍。
Suppression subtractive hybridization was used to investigate the response of turbot, Psetta maxima (L.), to Vibrio harveyi, by using a cDNA library constructed from artificially infected turbot kidney and spleen mRNA. Forty-nine expressed sequence tags were obtained. Several immune system genes were identified, including a major histocompatibility complex (MHC) class Ia gene and a heat shock protein 70 gene. Some signalling molecules were also present in the cDNA libraries, including src-family tyrosine kinase SCK, sgk-1 serine-threonine protein kinase and amyloid precursor-like protein 2. The full length of MHC class Ia cDNA was cloned from turbot cDNA by rapid amplification of cDNA ends polymerase chain reaction. The nucleotide sequence of turbot MHC class Ia has been submitted to GenBank with accession number EF032639. The turbot MHC class Ia cDNA has an open reading frame encoding 354 amino acids, and the deduced amino acid sequence of turbot MHC class Ia has 68%, 54%, 51%, 52%, 57%, 33%, 29% and 29% identities to those of olive flounder, medaka, rainbow trout, Atlantic cod, tiger puffer, chicken, mouse and human, respectively. Quantitative reverse transcriptase-PCR was performed for the MHC class Ia gene, and it was revealed that the expression level of the MHC class Ia gene in V. harveyi-challenged turbot increased to fourfold that of the controls.