番鸭IFN-α基因的克隆与序列分析

Cloning and Sequencing of the Muscovy Duck IFN1 Gene

从经免疫的番鸭外周血分离淋巴细胞 ,经PHA刺激培养后提取总RNA ,采用RT PCR的方法 ,按照GenBank中序列号为X84 76 4鸭I型IFN(DIFN1)基因设计引物 ,成功扩增出番鸭IFN基因 ,命名为MDIFN α ,包含编码MDIFN α成熟蛋白全部核苷酸序列 .DIFN1开放阅读框架 (ORF)有 5 76个核苷酸 ,编码 191个氨基酸 ,其中切除信号肽的IFN α为成熟DIFN1,共有 4 86个核苷酸组成 ,编码 16 1个氨基酸 .MDIFN α(含有不完整的信号肽 )与X84 76 4的核苷酸同源性为 97 7% ,推导氨基酸同源性为 95 7% .结果显示 ,MDIFN α可能为鸭I型IFN一个新的亚型 .

Using primers based on entry No.X84764 from GenBank, the muscovy duck IFN1 gene (named MDIFN α) containing the nucleotide sequence of the mature protein was cloned from total RNA derived from PHA stimulated cultured lymphocyte of immunized muscovy duck by reverse transcription polymerase chain reaction (RT PCR). The open reading frame (ORF) of DIFN1 consisted of 576 nucleotides, encoding 191 amino acids. After excising the signal peptide, mature DIFN1 consisted of 486 nucleotides, encoding 191 amino acids. The homology of MDIFN α(lacking signal peptide)nucleotide and deduced amino acid sequences to No.X84764 was 97.7% and 95.7% respectively. The result showed that MDIFN α might be a new IFN subtype of duck type I interferon.