中国旱獭α干扰素家族基因的克隆及序列分析

Cloning and sequencing of interferon alpha family gene of Chinese marmot

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 investigate the function of interferon alpha（IFN-α） in the Chinese marmot （ Marmota himalayana）, an animal model of hepatitis B. Methods Plasmids harboring Chinese marmot interferon alpha gene （IFNA） with different genotype were generated with molecular cloning technology. Sequence was sequenced, subtyped and put up homologous analysis and phylogenetic analysis. Results The results indicate that Chinese marmot IFN-α gene is a multigenic family from 8 IFNA functional sequences and 4 pseudogene sequences, The overall identity of the amino acid sequence among the members of the Chinese marmot IFN-α family is 85 %, and the identity with the IFN-α farrily from other species such as mice and humans is 53%. Conclusion We creat a tool to test the antiviral effect of new forms of IFN-α delivery and to improve the efficacy of current therapies in Chinese marmot HBV model.