摘要
从性成熟的福建黄兔母兔脑垂体中提取总RNA,经RT-PCR扩增目的片段,获得长为1 275bp的黄兔卵泡抑制素βA亚基cDNA片段。将扩增的片段与pMD18-T载体连接,转化至DH5α感受态细胞中。随机挑选阳性重组子进行鉴定、测序,将测序结果与包括人、大猩猩、奶牛、绵羊、小鼠在内的多种哺乳动物基因序列进行同源性比对分析,相似度在85%以上,同时遗传进化分析结果表明兔形目的INHBA基因与灵长目动物的亲缘性相近。黄兔INHBA基因cDNA序列编码424个氨基酸,整个蛋白包括1段20个氨基酸的信号肽和404个氨基酸的成熟肽,该蛋白还同时存在2个结构域,分别为TGFβ和TGFβ前体肽结构域。
The total RNA obtained from the pituitary of Fujian yellow rabbit was used to amplify the cDNA for the precursor of Fujian yellow rabbit inhibin βA subunit by RT-PCR, and the opening reading frame of 1 275 bps was acquired. The fragments amplified were ligated with pMD18-T vector and trans- formed into DHSa competent cells. The positive clones were verified, sequenced and submitted to Gen- Bank, with accession. KC831577. The sequence result was compared with homologous sequences from mammalian animals including human, cow, sheep, and mouse. Sequence alignment showed that ranks of their similarities were above 95%, and meanwhile, the results of genetic evolution analysis of the gene showed a fairly high degree of homology between rabbits and primates. The cDNA sequences of INHBA of Fujian yellow rabbit encodesd 424 amino acid,including a signal peptide containing 20 amino acid and a mature peptide containing 404 amino acid, and meanwhile the peptide were composed by structural do- mains of TGFβand TGFβ propeptide, respectively.
出处
《家畜生态学报》
北大核心
2013年第9期17-21,共5页
Journal of Domestic Animal Ecology
基金
国家兔产业体系福州综合试验站(CARS-44-E-11)
福建省自然科学基金(2012J05049)
福建省农业科学院博士科研启动基金(2011BS-2)