构建稳定表达JSRV受体Hyal-2小鼠肺上皮细胞系  

Construction of a TC- 1 cell line stably expressing hyaluronidase-2 as a receptor for jaagsiekte seep retrovirus

作　　者：孙丽红[1,2] 骆爽[1,2] 张德纯 斯日古楞[1,2] 么宏强[1,2] 

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]农业部动物疾病临床诊疗技术重点实验室,呼和浩特010018 [3]辽宁省沈阳市辽中县肖寨门动监所,沈阳110207

出　　处：《黑龙江畜牧兽医》2015年第12期211-213,318,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31260593)

摘　　要：为了构建稳定表达绵羊肺腺瘤病毒(JSRV)受体透明质酸酶-2(Hyal-2)蛋白的小鼠肺上皮细胞系,试验采用将重组质粒p DsRed-monomer-N1-Hyal-2转染小鼠肺上皮细胞后,用G418对转染后的细胞进行筛选、纯化,通过对不同培养代次的细胞系进行倒置荧光显微镜观察、RT-PCR、基因测序、Western-blot等方法加以验证。结果表明:试验成功构建了JSRV受体Hyal-2蛋白小鼠肺上皮细胞系,在传至20代后Hyal-2仍在该细胞系中稳定表达。说明该细胞系的成功建立,为研究绵羊Hyal-2对JSRV转化目的细胞能力的影响提供了一种体外试验平台。To construct a mouse lung epithelial cell line stably expressing the protein from hyaluronidase -2 （Hyal -2 ） protein a receptor for jaagsiekte sheep retrovirus （JSRV）, a recombinant plasmid pDsRed -monomer- N1 - Hyal -2 was used to transfeet the TC - 1 mouLse lung epithelial cells. The transfected cells were screened and purified using G418. The TC - 1 cell lines from different culture generations were veri- fied using a lot of methods including the observation of inverted fluorescence microscope, RT - PCR, gene sequencing, Western - blot, and so on. The results showed that a TC - 1 cell line expressing the protein from Hyal - 2 as a receptor for JSRV was successfully established, and the Hyal - 2 after transmitted to the 20 generations was stably expressed in the TC - 1 cell line. The successful establishment of the cell line provid- ed a kind of in vitro test platform for studying the effect of Hyal - 2 on the activity of JSRV transforming the target cell.

关 键 词：绵羊肺腺瘤病毒(JSRV) 透明质酸酶-2(Hyal-2) 受体 转染 小鼠肺上皮细胞系 

分 类 号：S858.26[农业科学—临床兽医学] S852.659.1[农业科学—兽医学]