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为分析鳜鱼(Siniperca chuatsi)生肌调节因子基因(mrf4)的特征,阐明其在鳜鱼不同组织以及胚胎发育不同时期的表达规律,用反转录PCR法扩增鳜鱼mrf4的cDNA序列,并将其连入克隆载体,进行测序、分析;用荧光定量PCR检测mrf4基因在不同组织及胚胎发育不同阶段的表达情况。结果:克隆的鳜鱼mrf4 cDNA全长为726 bp,编码的蛋白质含241个氨基酸,具有生肌调节因子的典型的碱性螺旋–环–螺旋结构;mrf4在成体鳜鱼白肌、红肌、心脏、脑以及不同发育阶段的胚胎中均有表达,在红肌中的表达量显著高于在其他组织中的表达量(P<0.05);在胚胎发育阶段的原肠期、尾芽期、肌肉效应期,mrf4的表达量呈递增趋势,而在胚胎发育的心搏期、血液循环期、仔鱼期,mrf4的表达量呈递减趋势,肌肉效应期mrf4的表达量明显高于胚胎发育的其他阶段的mrf4表达量(P<0.05)。
In order to analyze the characteristics of mrf4 in Siniperca chuatsi, and to elucidate the expression pattern of mrf4 in different tissues and embryos of anchovy, the cDNA sequence of mrf4 was amplified by reverse transcription polymerase chain reaction (RT-PCR) And linked into the cloning vector for sequencing and analysis. The expression of mrf4 gene in different tissues and different stages of embryonic development was detected by real-time PCR. Results: The cloned mrf4 cDNA was 726 bp in length and encoded a protein of 241 amino acids with a typical basic helix-loop-helix structure of myogenic modulators. The mrf4 gene was expressed in adult white sole muscle, red muscle, heart , Brain and embryos at different developmental stages. The expression levels in red muscle were significantly higher than those in other tissues (P <0.05). In the embryonic stage of the intestine, tail bud and muscle In the effector stage, the expression of mrf4 showed an increasing trend. However, the expression of mrf4 showed a decreasing trend during embryonic development in cardiac, blood circulation and larval stages. The expression level of mrf4 in muscle-responsiveness was significantly higher than that in other stages of embryonic development Of mrf4 expression (P <0.05).