表达超抗原sEA基因的溶瘤腺病毒载体的构建与鉴定(英文)
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范涛 郝林 贺厚光 梁清 董秉政 胡建鹏 韩从辉 徐州市中心医院泌尿外科;
【摘要】目的:构建表达超抗原SEA基因的溶瘤腺病毒载体并鉴定。方法:采用PCR技术,从产SEA的葡萄球菌标准菌株ATCC13565基因组DNA中获得SEA全长基因序列,酶切后克隆入pCA13质粒,构建重组病毒质粒pCA13-SEA。将鉴定正确的pCA13-SEA与含有腺病毒右臂的质粒pBHGE3通过Lipofectamine 2000共转染HEK293细胞,经同源重组产生重组腺病毒Ad-SEA。Ad-SEA在293细胞中大量扩增并通过氯化铯密度梯度离心法纯化、测定其滴度。结果:经PCR扩增、酶切鉴定、序列测定证实,SEA基因成功克隆到溶瘤腺病毒载体中,可实现SEA基因的表达。结论:成功构建了表达超抗原SEA基因的溶瘤腺病毒载体,为进一步研究该病毒对膀胱肿瘤靶向治疗的作用奠定了基础。
【关键词】 溶瘤腺病毒 超抗原SEA 肿瘤 载体
【基金】National Natural Science Foundation(30973443) Science and technology project of Xuzhou(XM08C055)~~
【分类号】R737.14
It is crucial that to select a proper vector for introducing thetarget gene to the target organ in the gene therapy of tumor.Manystudy mainly research the virus vectors,including the retrovirusvector,adenoviral vector,and adeno-associated virus(AAV)vec-t
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It is crucial that to select a proper vector for introducing thetarget gene to the target organ in the gene therapy of tumor. Manystudy mainly research the virus vectors, including the retrovirusvector, adenoviral vector, and adeno-associated virus (AAV) vec-tor, among which the adeno-associated virus vector is the mostpromising. Recently, a new virus vector, replication-selective on-colytic adenovirus (RSOAds), has attracted more attention. As akind of special virus gene therapy vector with the anti-rumor effect,it has the capabilities of selectively infecting the tumor cells andconducting large amplification and cracking the tumor cells.
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