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目的克隆中国旱獭α干扰素(IFNα)家族基因,以期利用克隆的中国旱獭家族IFNα在动物模型上探索慢性乙型肝炎有效的干扰素治疗方案和策略。方法利用分子克隆技术对中国旱獭IFNα家族基因进行克隆,并对所克隆的系列基因进行测序、分型、系统发生树分析、同源性比较及理化特性分析。结果从112个中国旱獭IFNα基因克隆中获得18个独立的不重复序列,其中的14个序列来自4次以上相对独立的PCR产物,将它们分为14个基因亚型,其中8个是功能基因亚型,6个是假基因亚型。中国旱獭IFNα各基因亚型之间在核苷酸水平和氨基酸水平有很高的同源性,平均分别为93%和85%,前体蛋白N端含23个氨基酸的疏水性信号肽。结论中国旱獭IFNα家族至少有14个基因亚型,这些基因的克隆可能应用于中国旱獭HBV动物感染模型,进行干扰素基因治疗和研究干扰素治疗策略。
OBJECTIVE: To clone IFNα family genes from Chinese marmota and explore the effective interferon therapeutic strategies and strategies for chronic hepatitis B in animal models using cloned Chinese marmot family IFNα. Methods The gene of IFNα family of Chinese marmot was cloned by molecular cloning technique. The cloned genes were sequenced, sequenced, phylogenetic tree analysis, homology comparison and physical and chemical properties analysis. Results Eighteen independent non-repetitive sequences were obtained from 112 Chinese mound IFNα gene clones. Among them, 14 sequences were from more than 4 independent PCR products, which were divided into 14 subtypes, of which 8 were functional genes Subtypes, 6 are pseudogene subtypes. The genotypes of IFNα in Chinese marmot had high homology at the nucleotide and amino acid levels, averaging 93% and 85% respectively, and a 23 amino acid hydrophobic signal peptide at the N-terminus of the precursor protein. Conclusion At least 14 subgenotypes of IFNα family of Chinese marmot are available. The cloning of these genes may be applied to the animal model of HBV infection in Chinese marmot, interferon gene therapy and interferon therapy strategy.