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[目的]获得松江鲈β-肌动蛋白基因cDNA全长序列,并检测松江鲈β-肌动蛋白在组织中的表达。[方法]以松江鲈肌肉总RNA为模板,采用RT-PCR、5′-RACE和3′-RACE的方法扩增β-肌动蛋白基因cDNA片段,用RT-PCR方法检测组织表达。[结果]获得了松江鲈β-肌动蛋白基因cDNA的3个片段,测序后拼接得到1905bp全长cDNA序列,其包含了1128个核苷酸的开放性阅读框,翻译编码375个氨基酸。核苷酸和氨基酸同源性分析发现,松江鲈β-肌动蛋白基因序列与点带石斑鱼、军曹鱼、红鲷鱼等同源性相对较高,与哺乳动物和鸟类同源性相对较低;系统发育分析表明,松江鲈β-肌动蛋白与点带石斑鱼关系最近。RT-PCR分析表明,该基因在检测的肌肉、肝脏、肠和脑4种组织均有表达。[结论]首次得到了β-肌动蛋白基因cDNA全长序列,并证明了松江鲈的β-肌动蛋白基因非常保守。
[Objective] The aim of the study was to obtain the full-length cDNA sequence of β-actin gene of Song bass and to determine the expression of β-actin in the tissue of the Japanese sea bass. [Method] The cDNA of β-actin gene was amplified by RT-PCR, 5’-RACE and 3’-RACE using the total RNA of P. japonicus as template. The expression of β-actin gene was detected by RT-PCR. [Result] Three fragments of cDNA of β-actin gene were obtained. After sequencing, the full-length cDNA sequence of 1905bp was obtained, which contained an open reading frame of 1128 nucleotides and translated 375 amino acids. Nucleotide and amino acid homology analysis showed that the β-actin gene sequence of P. japonicus showed relatively high homology with grouper, cobia, red snapper, and homology with mammals and birds Lower; Phylogenetic analysis showed that β-actin was the most closely related to grouper. RT-PCR analysis showed that the gene was expressed in the four tissues of muscle, liver, intestine and brain. [Conclusion] The full-length cDNA sequence of β-actin gene was obtained for the first time and it was proved that the β-actin gene of Song boa was very conservative.