鼠源巨噬细胞RAW264.7中TNF-α基因的克隆及酶切鉴定  

The source of the TNF alpha gene cloning and enzyme identification in the rat macrophage RAW264.7

作　　者：赵义龙[1] 黄金凤 赵金香[3] 矫继峰[3] Zhao Yilong Huang Jin-feng Zhao Jin-xiang JiaoJiFeng(xinjiang agricultural vocational and technical college xinjiang changji 831100 xinjiang Wujiaqu xin bao agricultural science and technology development company xinjiang wujiaqu 831100 YingKou institute liaoning yingkou 115014)

机构地区：[1]新疆农业职业技术学院,新疆昌吉831100 [2]新疆五家渠鑫宝农业科技开发有限公司,新疆五家渠831300 [3]营口理工学院,辽宁营口115014

出　　处：《广东畜牧兽医科技》2017年第3期42-44,共3页Guangdong Journal of Animal and Veterinary Science

基　　金：2016年度新疆农业职业技术学院资助项目(XJNZYKJ2016030)

摘　　要：TNF-α能诱导单核巨噬细胞系统的前体细胞分化,是机体非特异性免疫检测中指示性细胞因子。本研究根据Genbank NM-013693上登录的小鼠肿瘤坏死因子-α(TNF-α)序列对经过氨基多糖纳米微粒处理过的小鼠RAW264.7细胞进行RNA的提取,且RT-PCR扩增TNF-α基因。将此片段克隆到PUCm-T载体中,经菌落PCR鉴定和DNA序列测定、酶切鉴定分析验证,证实所克隆序列为TNF-α基因。序列分析表明,该基因与Genbank中发表序列的同源性达到100%。the TNF alpha can induce progenitor cell differentiation of mononuclear macrophage system, is the body＇s nonspecific immune detection indicative of cytokines. This study based on Genbank NM-013693 logged in mice tumor necrosis factor alpha（TNF alpha） sequence of mice that had not been treated with amino polysaccharide nanoparticles RAW264.7 cell to the extraction of RNA, and RT-PCR amplification TNF alpha gene. This fragment cloned in PUCm-T carrier, DNA sequencing, enzyme by colony PCR identification and appraisal analysis, confirm the sequence of TNF alpha gene cloning. Sequence analysis showed that the gene in Genbank published sequence homology of 100%.

关 键 词：RAW264.7 TNF-Α PCR 克隆 酶切鉴定 序列分析 

分 类 号：Q78[生物学—分子生物学]