牛γ-干扰素基因的克隆及序列分析

Cloning and Sequence Analysis of Bovine Interferon-gamma Gene

根据GenBank中已公布的牛γ-干扰素(BovIFN-γ)基因序列设计引物,应用一步法逆转录—聚合酶链式反应(RT-PCR)技术,以从牛外周血淋巴细胞提取的总RNA为模板,扩增出γ-干扰素成熟肽基因片断,并对其进行了克隆、测序及序列分析。结果表明,试验所设计引物可以成功用于扩增牛γ-干扰素成熟肽基因片段,所扩增序列与GenBank中已有序列同源性达99.77%。

Based on the published nucleotide sequence of bovine interferon-gamma （BovIFN-γ） gene, a pair of RT-PCR primers were designed and synthesized. The mature peptide fragments of γ-interferon gene was amplified using reverse transcription polymerase chain reaction （RT-PCR）from total RNA of bovine peripheral blood lymphocytes, and then carried out cloning, sequencing and sequence analysis. The results showed that the mature peptide of BovIFN-γ gene can be successfully amplified by primers designed. Additionally, the homology between the amplified gene fragment sequence and the relevant sequence in GenBank can reach up to 99.77 %.