北京鸭干扰素-α基因的分子克隆与表达

Molecular cloning and expression of Peking duck IFN-α gene

采用 PCR法从北京鸭 (Anas platyrhynchosdoestica L innaeus)基因组中克隆了其干扰素 - α基因 (BJ- Du IFN- α)。克隆片段长 4 86 bp,编码 1 6 1个 Du IFN-α成熟肽。与已报道的北京鸭干扰素 -α基因相比 ,BJ- Du IFN-α在 2 85位发生了同义变异。将 BJ-Du IFN-α插入原核表达载体 p QE3 0 ,在 E.Coli JM1 0 9工程菌中表达了重组蛋白。经 SDS- PAGE、Western Blot和抗病毒活性测定 ,表达产物占菌体总蛋白的 2 3 %,为重组鸭干扰素 -α(r Du IFN-α)。 r Du IFN-α抗滤泡性口炎病毒 (VSV)活性高达 7.9× 1 0 5U /mg。

Peking Duck (Anas platyrhynchos doestica Linnaeus) Interferon-alpha gene (BJ-DuIFN-α) was cloned from the genomic DNA using PCR method. The cloned gene was 486 bp in length and encoded 161 animo acids. Comparison of BJ-DuIFN-α with DuIFN-α gene, only one substitution was found at 285 site (T-C). BJ-DuIFN-α was inserted into pQE30 expression vector, and the JM109/pQE30/BJ-DuIFN-α engineering bacterial proteins indentified by SDS-PAGE and Western blot analysis. The anti-virus bioactivity was high up to 7.9×10~5 U/mg.