团头鲂MSTN基因cDNA结构、表达及过表达对胚胎发育的影响

发布时间：2018-12-30 21:00

【摘要】：研究通过c DNA末端快速扩增法(RACE)克隆得到团头鲂生长抑制素(MSTN)基因的c DNA全长并分析了MSTN基因在团头鲂胚胎、成鱼组织中表达以及MSTN基因在胚胎中过表达情况。结果表明团头鲂MSTN基因的c DNA全长为2187 bp,ORF(开放阅读框)大小为1128 bp,编码376个氨基酸。组织逆转录PCR(RT-PCR)结果显示,MSTN基因在肌肉、脑和精巢组织中大量表达,肝脏、脾脏和卵巢组织中的少量表达,肠、腮、心、眼和肾组织中的微量表达。胚胎逆转录PCR(RT-PCR)结果显示,在0—44 hpf胚胎发育阶段,MSTN基因表达量较低;而在48—52 hpf胚胎发育阶段,MSTN基因表达量逐渐升高。整胚原位杂交(WISH)结果显示,胚胎发育的16 hpf时期MSTN基因主要在脊索中表达,胚胎发育的28 hpf和55 hpf时期MSTN基因在脑中表达。MSTN基因过表达结果显示,胚胎在体节发生期出现前-后轴拉长,背-腹轴变短;脊索发生扭曲,强烈抑制体节发育而导致不分化等现象。研究为后续团头鲂MSTN基因的功能研究及团头鲂分子育种提供相关参考依据。
[Abstract]:The full length of c DNA of (MSTN) gene was cloned by rapid amplification of c DNA terminal (RACE). The expression of MSTN gene in embryo and adult fish tissues and overexpression of MSTN gene in embryo were analyzed. The results showed that the full length of c DNA of MSTN gene was 2187 bp,ORF (open reading frame) and the size of c DNA was 1128 bp, encoding 376 amino acids. Tissue reverse transcription PCR (RT-PCR) showed that MSTN gene was highly expressed in muscle, brain and spermary tissues, a small amount in liver, spleen and ovary, and in intestine, cheek, heart, eye and kidney. The results of embryo reverse transcription PCR (RT-PCR) showed that the expression of MSTN gene was lower in 0-44 hpf embryo development stage, but increased gradually in 48-52 hpf embryo development stage. The results of in situ hybridization (WISH) showed that the MSTN gene was mainly expressed in the notochord at 16 hpf of embryonic development, MSTN gene was expressed in the brain at 28 hpf and 55 hpf in embryonic development, and MSTN gene was overexpressed in the brain. In the ganglionogenesis stage, the pre- posterior axis was elongated, and the dorsol-ventral axis became shorter. The notochord is distorted, strongly inhibits the development of somatic ganglia and leads to indifferentiation. The study provides a reference for the functional study of MSTN gene and molecular breeding of bream.
【作者单位】：上海海洋大学农业部淡水水产种质资源重点实验室;
【基金】：国家科技支撑计划(2012BAD26B00) 国家自然科学基金(31272633和31201760)资助~~
【分类号】：S917.4

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