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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2021, Vol. 47 Issue (4): 534-542    DOI: 10.3785/j.issn.1008-9209.2020.10.222
动物科学与动物医学     
不同品系豚鼠MHCⅠ类基因的多态性及差异表达
卫振1(),何珂2,洪胜辉1,刘迪文1()
1.浙江大学实验动物中心，杭州 310058
2.浙江农林大学动物科技学院和动物医学院，杭州 311300
Polymorphism and differential expression of MHCclassⅠgenes between different strains of guinea pig
Zhen WEI1(),Ke HE2,Shenghui HONG1,Diwen LIU1()
1.Laboratory Animal Center, Zhejiang University, Hangzhou 310058, China
2.College of Animal Science and Technology/College of Veterinary Medicine, Zhejiang A&F University, Hangzhou 311300, China
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摘要：

由前期的研究可知，Zmu-1∶DHP远交系豚鼠对口蹄疫病毒的敏感性为100%，和其他DHP品系相比存在显著差异。本研究利用不同品系豚鼠（Zmu-1∶DHP远交系、Zmu-2∶DHP远交系和DHP品系），提取其脾脏RNA，转录为cDNA后进行聚合酶链式反应扩增，扩增产物经过高通量测序，分析豚鼠MHCⅠ类基因结构、单倍型及多态性信息，以检测各品系间该类基因的表达是否存在差异。结果表明：本研究系统地得到豚鼠的8条MHCⅠ类基因单倍型序列，其中部分单倍型在外显子3存在23个氨基酸缺失，但依旧存在一定程度的表达；部分品系间单倍型表达测序片段（reads）频率差异显著（P＜0.05），推测MHCⅠ类基因单倍型表达差异可能和不同品系的免疫能力差异有关。本研究为豚鼠品系应用于疾病模型及疫苗开发奠定了理论基础。
关键词： 豚鼠;  Zmu品系;  MHCⅠ类基因    
Abstract:

Guinea pig is a laboratory animal model to study the illness and stimulation of infection. The previous studies showed that the sensitivity of Zmu-1∶DHP outbred guinea pig to foot-and-mouth disease virus was 100%, which was significantly different from DHP (parental strain). This study took three strains of guinea pigs (Zmu-1∶DHP outbred line, Zmu-2∶DHP outbred line and DHP strain) as the experimental groups. The RNA extracted from spleen of different strains of guinea pigs was transcribed into cDNA and amplified by polymerase chain reaction. The amplified products were sequenced by high-throughput sequencing to analyze the structure, haplotype and polymorphism of MHC classⅠgene in guinea pigs, so as to detect whether there were differences in the expression of MHC classⅠgene among different strains. The results showed that eight MHC classⅠsequences in guinea pig were found and some of these haplotypes had 23 amino acids deletion in exon 3. There were significant differences in the frequencies of haplotype expression reads among some strains (P＜0.05), which suggested that the difference of MHCclassⅠhaplotype expression might be related to the difference of immune ability among different strains. The research provides a theoretical basis for the application of guinea pig strains in disease model and vaccine development.
Key words: guinea pig    Zmu strain    MHC classⅠgene
收稿日期: 2020-10-22 出版日期: 2021-08-25
CLC:  S 813  
基金资助: 卫生部科学研究基金(98-2-823)
通讯作者: 刘迪文     E-mail: lac@zju.edu.cn;liudiwen2004@163.com
作者简介: 卫振（https://orcid.org/0000-0001-8824-8411），E-mail：lac@zju.edu.cn
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卫振
何珂
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引用本文:

卫振,何珂,洪胜辉,刘迪文. 不同品系豚鼠MHCⅠ类基因的多态性及差异表达[J]. 浙江大学学报（农业与生命科学版）, 2021, 47(4): 534-542.

Zhen WEI,Ke HE,Shenghui HONG,Diwen LIU. Polymorphism and differential expression of MHCclassⅠgenes between different strains of guinea pig. Journal of Zhejiang University (Agriculture and Life Sciences), 2021, 47(4): 534-542.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2020.10.222    或    http://www.zjujournals.com/agr/CN/Y2021/V47/I4/534

图1  本研究所用的PCR引物示意图

单倍型

Haplotype

BWMB vs WB vs MW vs M
均值Mean

标准差

Standard deviation

均值

Mean

标准差

Standard deviation

均值

Mean

标准差

Standard deviation

Capo-ⅠA*010.0820.0060.0640.0150.0820.010**NS**
Capo-ⅠA*020.1400.0080.1390.0100.1370.005NSNSNS
Capo-ⅠA*030.0310.0060.0250.0060.0300.006*NSNS
Capo-ⅠA*040.1540.0150.1650.0140.1490.011NSNS**
Capo-ⅠA*050.0950.0130.1200.0180.0960.013**NS**
Capo-ⅠA*060.3690.0220.3700.0280.3850.006NS*NS
Capo-ⅠA*070.0830.0040.0740.0110.0800.007*NSNS
Capo-ⅠA*080.0450.0090.0430.0160.0400.008NSNSNS
表1  3个品系间MHCⅠ类基因单倍型频率分析及比较
图2  豚鼠MHCⅠ类基因单倍型多态性分析图中蓝色、红色和绿色区域分别表示α1、α2和Capo-ⅠA*06的缺失区域；“＋”表示抗原结合域，参考序列为HLA（D38525.1）。
图3  单倍型Capo-ⅠA*01和Capo-ⅠA*06的蛋白质三维结构预测图图中红色表示部分单倍型中的缺失区域，粉色表示二硫键结合区域。
图4  豚鼠MHCⅠ类序列系统发育树带标记的进化支已被调整，不与图示进化距离一致。
图5  部分哺乳物种MHCⅠ类序列的系统发育树
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