鸡SAMD9L基因真核表达载体构建及其对ALV-J病毒复制的影响

被引：7
作者：
陈世豪 [1 ]
潘诗雨 [1 ]
赵睿涵 [1 ,2 ]
吴挺 [1 ]
机构：
[1] 扬州大学表观遗传学与表观基因组学研究所
[2] 扬州大学兽医学院
来源：
扬州大学学报(农业与生命科学版) | 2021年 / 42卷 / 06期
基金：
中国博士后科学基金;
关键词：
鸡; SAMD9L; 生物信息学分析; 真核表达载体; ALV-J复制;
D O I：
10.16872/j.cnki.1671-4652.2021.06.009
中图分类号：
S858.31 [鸡];
学科分类号：
摘要：
为分析鸡无菌α基序域蛋白9样(SAMD9L)蛋白的生物学特性,构建SAMD9L基因真核表达载体,并探究其对J亚型禽白血病病毒(ALV-J)复制的影响。利用ProtParam、Uniport、TMHMM和String等在线网站分析SAMD9L序列,获得亲疏水性、保守结构域、信号肽表达及互作蛋白等生物信息。构建携带His标签的SAMD9L重组表达载体并进行测序鉴定。采用qRT-PCR技术检测ALV-J感染对SAMD9L表达水平的影响;采用Western blot技术检测过表达SAMD9L对ALV-J复制的影响。结果表明:鸡SAMD9L无信号肽表达,无跨膜结构域,含有无菌α基序超家族保守结域(55—249位氨基酸),其互作蛋白预测为MX1、DDX60、ISG12-2和IFIT5等天然免疫抗病毒蛋白质。ALV-J感染36 h后显著诱导SAMD9L表达(P<0.001),而过表达SAMD9L则显著抑制ALV-J在DF-1细胞中的复制,推测鸡SAMD9L具有抗ALV-J病毒的能力。该研究成功构建了鸡SAMD9L基因真核表达载体,通过生物信息学分析了蛋白质特性,并初步探究SAMD9L与ALV-J感染之间的关系,为进一步阐明鸡SAMD9L的生物学功能及其抗J亚型禽白血病的分子机制奠定了基础。
引用
收藏
页码：54 / 59
页数：6
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