| 茅莉娜,冯娟,李玉谷,侯月娥,郭志勋,许海东.2010.军曹鱼MHC-Ⅱβ基因全长cDNA的克隆与组织表达分析.动物学杂志,45(4):118-128. |
| 军曹鱼MHC-Ⅱβ基因全长cDNA的克隆与组织表达分析 |
| Full Length cDNA Cloning and Tissue Expression of Major Histocompatibility Complex (MHC)-Ⅱ β in Cobia (Rachycentron canadum) |
| 投稿时间：2010-01-08 修订日期：2010-05-20 |
| DOI： |
| 中文关键词: 军曹鱼 MHC-Ⅱβ 全长cDNA 组织特异性表达 |
| 英文关键词:Cobia Rachycentron canadum MHC-Ⅱβ Full length cDNA mRNA expression |
| 基金项目:中央级公益性科研院所基本科研业务费专项资金项目(No.2007ZD10) |
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| 中文摘要: |
| 采用同源克隆和末端快速扩增(RACE)的方法,得到1161bp的军曹鱼(Rachycentron canadum)MHC-Ⅱβ全长cDNA片段。该序列包括20bp的5′末端非编码区(UTR),394bp的3′UTR及747bp的开放阅读框(ORF),编码248个氨基酸,预测其蛋白质分子量约为27.99ku,等电点为6.21。通过构建MHC-Ⅱβ氨基酸序列的系统进化树,并进行氨基酸相似性比对,结果表明,军曹鱼和已知鱼类、鼠(Musmusculus)、鸡(Gallus gallus)及人类(Homo sapiens)MHC-Ⅱβ氨基酸的同源性在28.5%～77.5%之间。所推测的蛋白序列具有一些重要的特征,包括:前导肽、β1、β2和CP/TM/CYT区,保守的半胱氨酸等。Real-time PCR检测结果显示,MHC-Ⅱβ基因在正常军曹鱼组织中均表达,但表达量在各个组织中不同,其中,头肾表达较强,鳃、脾、肠表达程度中等,在心、脑、肌肉中表达较弱。 |
| 英文摘要: |
| The homology cloning and RACE PCR were employed to clone the MHC-Ⅱ β gene from cobia (Rachycentron canadium). The full length cDNA of MHC-Ⅱβ comprises 1 161 bp with a 20 bp 5′ untranslated region (UTR),a 394 bp 3′ UTR and a 747 bp open reading frame (ORF) encoding a polypeptide of 248 amino acid residues with a predicted molecular weight of 27. 99 ku and theoretical isoelectric point of 6. 21. The putative protein showed homology varying from 28. 5% to 77. 5% when compared with other fishes,mouse(Mus musculus),chicken(Gallus gallus)and human (Homo sapiens). The protein sequence showed all the important features including leader peptide,β1,β2,CP /TM /CYT regions and conserved cysteines. The MHC-Ⅱ β expression was detected in all tested tissues with different expression levels. The high expression was detected in head kidney,moderate expression was detected in gill,spleen and intestine,while low expression was detected in heart,brain and muscle. |
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