| 摘要: |
| 利用RT-PCR和快速扩增cDNA末端(rapid amplification of cDNA ends, RACE)技术首次克隆了宽体沙鳅(Botia reevesae)β-肌动蛋白基因的cDNA全序列, 该序列全长为1795bp, 由长100bp的5'非翻译区(untranslated region, UTR)、570bp的3'非翻译区和1125bp的开放阅读框(open reading frame, ORF)组成, 编码375个氨基酸。宽体沙鳅β-actin氨基酸序列包含1个糖基化位点、9个N-豆寇酰化位点、3个actin信号位点等主要结构区域。PSI-BLAST比对表明, 宽体沙鳅β-actin氨基酸与真鲷、罗非鱼、虹鳟等鱼类同源性达99%。NJ法系统进化分析显示宽体沙鳅β-actin首先与鲢聚在一起, 然后与 鳡、尖头鱥等鱼类聚在一起。荧光定量PCR检测β-actin基因在宽体沙鳅脑、鳃、心脏、肝、胃等12个组织的表达无显著差异(P<0.05), 具有良好的稳定性。 |
| 关键词: 肌动蛋白 宽体沙鳅 cDNA 组织表达 |
| DOI：10.11693/hyhz201302021021 |
| 分类号: |
| 基金项目:四川省教育厅项目, 11ZB025 号; 四川省科技厅项目, 2011NZ0075 号; 国家科技支撑计划课题资助项目, 2012BAD25B03号; 公益性行业(农业)科研专项资助项目, 201203065 号; 内江师范学院大学生创新性实验计划项目, X201207 号 |
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| CLONING AND EXPRESSION ANALYSIS OF THE FULL-LENGTH cDNA SEQUENCE OF BOTIA REEVESAE β-ACTIN GENE |
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QIN Chuan-Jie1, CHEN Li-Qiao2, YUE Xing-Jian1, LI Er-Chao2, WANG Yong-Ming1, ZOU Yuan-Chao1, XIE Bi-Wen1, QI Ze-Min1
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1.School of Life Science, Neijiang Normal University;2.College of Life Science, East China Normal University
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| Abstract: |
| A 1795bp full-length cDNA sequence of β-actin gene from Botia reevesae was obtained with RT-PCR and rapid amplification of cDNA ends (RACE) technique. It consists of a 100bp 5' untranslated region (UTR), a 1125bp open reading frame (ORF) and a 570bp 3' UTR. The translated protein is composed of 375 amino acids. The putative domains include a N-glycosylation, nine N-myristoylation sites, and three actin signature in B. reevesae. Sequence comparison indicates that the β-actin deduced amino acid sequence of B. reevesae has an overall identity of 99%, 98% and 95% to that of Pagrus major,Oreochromis niloticus and Oncorhynchus mykiss, respectively. Alignment of deduced amino acid sequence to other species shows that the overall structure of β-actin is evolutionarily conserved. Phylogenetic analysis reveals that the B. reevesae β-actin is closely related to the β-actin in other fish. Quantitative PCR analysis shows that β-actin was equally expressed in the detected tissues. |
| Key words: actin gene Botia reevesae cDNA tissues expression |
