cDNA Cloning and Sequencing for Precursor of Fujian Yellow Rabbit Follicle Stimulating Hormone β Subunit
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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390bp的黄兔促卵泡素β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、奶牛、绵羊、水牛、小鼠在内的多种哺乳动物基因序列进行同源性分析, 同源性相似度72.2%96.5%, 同时遗传进化分析结果表明, 兔形目的FSHβ基因与灵长目动物的亲缘性更高。对福建黄兔与日本白兔已提交的FSHβ亚基的cDNA序列进行比较, 发现在81bp及219bp处存在C→T和G→A两处突变, 但未引起氨基酸变化。Abstract: The total RNA of the Fujian yellow rabbit′s pituitary was used to amplify the cDNA for the precursor of its follicle stimulating hormoneβsubunit (FSHβ) by means of RT-PCR.The opening reading frame of 390bp was obtained.The fragments amplified were ligated with pMD18-T vector and transformed into DH5αcompetent cells.Randomly selected positive clones were sequenced and submitted to GenBank for verification (accession number: KC784943) .It was compared with homologous sequences of the mammalian animals, including human, cow, sheep, buffalo, and mouse.The sequence alignment showed the similarities ranging from 72.2%to 96.5%.The genetic evolution analysis on the gene indicated a fairly high degree of homology of the rabbit with the primates.The cDNA sequences of the Fujian yellow rabbit′s FSHβwere also compared with those of the Japanese white rabbit′s.Nucleotide mutations at C81Tand G219Awere found, but not on amino acids.
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Key words:
- Fujian yellow rabbit /
- FSHβ /
- cloning /
- sequence analysis
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