腺病毒与慢病毒感染牛肌肉原代细胞的研究  

Study on Adenovirus and Lentivirus Infection of Bovine Muscle Primary Cells

作　　者：赵璐[1] 付长振 赵志东[2] 昝林森[2] 刘永峰[1] 

机构地区：[1]陕西师范大学食品工程与营养科学学院,陕西西安710062 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《安徽农业科学》2016年第4期147-150,185,共5页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(31372288);教育部高等学校博士学科点专项科研基金项目(20130202120008);陕西省青年科技新星项目(2014KJXX-51);中央高校基本科研业务费专项(GK201502008)

摘　　要：[目的]比较腺病毒和慢病毒侵染牛肌肉卫星细胞MOI值的差异,选择较优的侵染方式。[方法]从新生牛犊背最长肌中分离并纯化出牛肌肉卫星细胞,通过病毒侵染靶细胞的方法比较腺病毒和慢病毒对牛肌肉卫星细胞的侵染能力,确定最佳感染复数(Multiplicity of infection,MOI)。[结果]成功分离出牛肌肉卫星细胞,经过体外培养,可以用于细胞侵染研究;随着MOI值的增大,腺病毒和慢病毒侵染的细胞荧光强度均逐渐增强;当腺病毒侵染牛肌肉卫星细胞的MOI值为3 000,慢病毒侵染靶细胞的MOI值为300时,荧光强度均较强,细胞形态即将发生变化。相对而言,慢病毒较腺病毒更易侵染原代肌肉细胞,并且荧光强度较强,但侵染效率比较低。[结论]对于原代细胞,选择慢病毒侵染更为合适。该试验结果为应用腺病毒或慢病毒介导的功能基因在牛肌肉卫星细胞中的过表达或干扰沉默研究提供了参考资料。[Objective] The aim was to compare the difference of MOI values between the adenovirus and the lentivirus infected bovine muscle satellite cells,and to select the best method of infection. [Method] Bovine muscle satellite cells were isolated and purified from newborn calf of longissimus muscle,infection ability of adenovirus and lentivirus to bovine muscle satellite cells were compared to determine the optimal MOI. [Result] Bovine muscle satellite cells were successfully isolated,which can be used for the study of cell infection by cultured in vitro. It was found that,with the increase of MOI values fluorescence intensity gradually increased for infected cells,the MOI infection of bovine muscle satellite cells was 3 000 for adenovirus with strong fluorescence. While,lentivirus infection of target cells MOI increased to 300 having strong fluorescence,and morphology will be changed. In comparison,lentivirus more easily than adenovirus infected primary cells,and the fluorescence intensity was stronger,but the infection efficiency was relatively low. [Conclusion]It is appropriate to select lentivirus infection of primary cells. The result provides reference for research of cattle genes function via using denovirus or lentivirus genes mediated bovine muscle satellite cells express or disturbanced in silence.

关 键 词：牛肌肉卫星细胞 腺病毒 慢病毒 MOI 

分 类 号：S858.23[农业科学—临床兽医学]