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亚洲猫β-干扰素基因的克隆与分析
引用本文:刘宝山,朱冰,杨国丽,赵玉军.亚洲猫β-干扰素基因的克隆与分析[J].安徽农业科学,2008,36(3):904-905.
作者姓名:刘宝山  朱冰  杨国丽  赵玉军
作者单位:沈阳农业大学畜牧兽医学院,辽宁沈阳,110161
基金项目:沈阳农业大学青年教师科研基金项目(2006119).
摘    要:目的]为猫干扰素类生物制品的研究奠定基础。方法]根据NCBI上的猫IFN-β基因序列,设计1对特异引物。以从病死猫肝脏组织中提取的基因组DNA为模板,进行PCR扩增。将扩增产物回收后,与pUCm-T载体连接,并转化到大肠杆菌DH5α感受态细胞。通过PCR和双酶切对获得的质粒DNA进行鉴定后,进行序列分析和核苷酸、氨基酸序列同源性的比较。结果]经PCR扩增,可获得一条561 bp的特异性条带。猫IFN-β基因已成功重组到载体中,获得阳性克隆。序列分析结果表明,亚洲猫IFN-β基因长561 bp,编码186个氨基酸,与已报道的猫IFN-β基因同源性达99.82%,而氨基酸序列未改变。结论]该研究成功克隆了猫IFN-β基因,为进一步利用基因工程技术生产重组猫IFN-β基因及其生物学功能的研究奠定了基础。

关 键 词:β-干扰素  亚洲猫  克隆  分析
文章编号:0517-6611(2008)03-00904-02
收稿时间:2007-09-14
修稿时间:2007年9月14日

Cloning and Analysis of Interferon-β Gene in Asian Cat
LIU Bao-shan et al.Cloning and Analysis of Interferon-β Gene in Asian Cat[J].Journal of Anhui Agricultural Sciences,2008,36(3):904-905.
Authors:LIU Bao-shan
Abstract:Objective] The aim of the research was to lay the foundation for studying the biological products of cats such as interferon(IFN).Method] A pair of specific primers were designed according to the sequences of feline IFNβ gene on NCBI website.The genomic DNA extracted from the liver tissues of the diseased and dead cats was taken as template for PCR amplification.After being reclaimed,the amplified products were connected with pUCm-T vector and transformed into DH5α competent cells of Escherichia coli.After the obtained plasmid DNA was identified by PCR and double-enzyme digestion,the sequence analysis and the homology comparison of the nucleotides and amino acids were made.Result] A specific band of 561 bp was obtained from PCR amplification.Feline IFN-β gene was successfully recombined into the vector and the positive clone was obtained.The results of sequence analysis showed that IFN-β gene of Asian cat was at the length of 561 bp and it encoded 186 amino acids.Its homology with the reported feline IFN-β gene reached 99.82 % and the sequences of amino acids didn't change.Conclusion] This research successfully cloned feline IFN-β gene,and laid the foundation for further producing the recombinant feline IFN-β gene by gene engineering technology and studying its biological functions.
Keywords:Interferon-β  Asian cat  Cloning  Analysis
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