CAJ | 학술논문

　　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、尖头等鱼类聚在一起。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。鳡
　　이용RT-PCR화쾌속확증cDNA말단(rapid amplification of cDNA ends, RACE)기술수차극륭료관체사추(Botia reevesae)β-기동단백기인적cDNA전서렬,해서렬전장위1795bp,유장100bp적5′비번역구(untranslated region, UTR)、570bp적3′비번역구화1125bp적개방열독광(open reading frame, ORF)조성,편마375개안기산。관체사추β-actin안기산서렬포함1개당기화위점、9개N-두구선화위점、3개actin신호위점등주요결구구역。PSI-BLAST비대표명,관체사추β-actin안기산여진조、라비어、홍준등어류동원성체99%。NJ법계통진화분석현시관체사추β-actin수선여련취재일기,연후여、첨두등어류취재일기。형광정량PCR검측β-actin기인재관체사추뇌、새、심장、간、위등12개조직적표체무현저차이(P<0.05),구유량호적은정성。감
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 in-dicates that theβ-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.