CAJ | 학술논문

为揭示原肌球蛋白基因在草鱼肌肉中的作用,利用 RT-PCR 和 RACE 技术克隆获得了草鱼原肌球蛋白基因 cDNA,并对该基因在普通草鱼和脆肉鲩不同组织中的表达情况进行研究分析。结果表明原肌球蛋白基因cDNA全长序列为1705 bp,包含387 bp的5′UTR序列,1307 bp的3′UTR序列和855 bp开放阅读框(ORF)。其ORF编码284个氨基酸。系统进化分析表明普通草鱼与斑马鱼、墨西哥脂鲤的原肌球蛋白基因核苷酸同源性分别是93％和87％,氨基酸同源性分别是96％和93％。在聚类上普通草鱼原肌球蛋白基因与其他鲤科鱼类同源性较高,表明亲缘关系最近,与传统分类相一致。Real time-PCR结果表明原肌球蛋白基因在所检测的普通草鱼和脆肉鲩7个组织中均有表达,原肌球蛋白基因在普通草鱼腹肌中表达最高,其次为前肠。原肌球蛋白基因在脆肉鲩腹肌中的表达低于普通草鱼,而脆肉鲩中肌肉、肝脏、肾脏、前肠、后肠中原肌球蛋白基因表达量大于普通草鱼相对应组织,但差异不显著。
위게시원기구단백기인재초어기육중적작용,이용 RT-PCR 화 RACE 기술극륭획득료초어원기구단백기인 cDNA,병대해기인재보통초어화취육혼불동조직중적표체정황진행연구분석。결과표명원기구단백기인cDNA전장서렬위1705 bp,포함387 bp적5′UTR서렬,1307 bp적3′UTR서렬화855 bp개방열독광(ORF)。기ORF편마284개안기산。계통진화분석표명보통초어여반마어、묵서가지리적원기구단백기인핵감산동원성분별시93％화87％,안기산동원성분별시96％화93％。재취류상보통초어원기구단백기인여기타리과어류동원성교고,표명친연관계최근,여전통분류상일치。Real time-PCR결과표명원기구단백기인재소검측적보통초어화취육혼7개조직중균유표체,원기구단백기인재보통초어복기중표체최고,기차위전장。원기구단백기인재취육혼복기중적표체저우보통초어,이취육혼중기육、간장、신장、전장、후장중원기구단백기인표체량대우보통초어상대응조직,단차이불현저。
To reveal the original role of TPM gene in grass carp muscle, using rapid amplification of cDNA ends (RACE), the full length of TPM cDNA sequence in grass carp is cloned, and the expression of different tissues in common grass carp and embrittlement of grass carp are analyzed by Real-time PCR. The sequence analysis shows that the TPM cDNA had 1 705 bp in length with a 3′ -untranslated region (1 307 bp), a 5′-untranslated region (387 bp), and a 855 bp open reading frame (ORF). Bioinformatics analysis predicts that the ORF of TPM4a cDNA encoded a protein of 284 amino acid residuals with a structure of protein tertiary model with 277 alpha helix and 7 random curl. The phylogenetic analysis shows that the amino acid sequence of common grass carp TPM possessed 96%and 93%identity with the TPMs of Daniorerio and Astyanax mexicanus, respectively. Real-time PCR results demonstrated that the abdominal muscle of common grass carp had the highest expression of TPM, and next the foregut. Comparing with embrittlement of grass carp, TPM shows lower expression inabdominal muscle of common grass carp, while the higher expression of embrittlement of grass carp tissues were muscle, liver, spleen, foregut and hindgut. The results indicate that TPM can be a new candidate target gene for investigating embrittlement mechanism of grass carp embrittlement and improving healthy level of culture.