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核心蛋白聚糖真核表达载体的构建及其在A549中的表达 被引量：1

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摘要 目的建立人核心蛋白聚糖(decorin,DCN)真核表达载体,并在A549细胞中进行表达,为进一步研究其抗肿瘤作用奠定基础。方法用PCR法扩增出DCN分子编码序列的cDNA全长,与pcDNA3.1(+)载体连接,构建真核表达载体pcDNA3.1-decorin,并转化到大肠杆菌JM109中扩增以获得重组载体,应用双酶切、PCR以及测序鉴定此重组载体;脂质体介导重组载体转染A549细胞,经G418筛选建立稳定转染细胞株,分别采用RT-PCR、免疫组化法和Western印迹法检测其表达。结果获得了约1 080 bp大小的特异性DNA片段;PCR产物与真核表达载体进行连接,经过双酶切、PCR以及测序鉴定证实分泌型DCN cDNA片段正确插入真核表达载体pcDNA3.1中。RT-PCR方法可见转染组mRNA表达明显增多,免疫组化和Western印迹方法可见转染组细胞DCN蛋白表达明显增高。结论成功构建了真核表达载体pcDNA3.1-decorin,建立了稳定转染DCN的A549细胞株。
作者 吕俊锋 张玉成 杜珍武 张桂珍
机构地区 吉林大学第二医院放射线科 吉林大学中日联谊医院中心实验室
出处 《中国老年学杂志》 CAS CSCD 北大核心 2011年第5期803-806,共4页 Chinese Journal of Gerontology
基金 吉林省发展与改革委员会计划资助项目(200621550)
关键词 核心蛋白聚糖 基因表达 转染 A549细胞
分类号 Q786 [生物学—分子生物学]
  • 相关文献

参考文献8

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二级参考文献24

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共引文献5

同被引文献11

  • 1Silvia Goldoni,Daniela G. Seidler,Jack Heath,Matteo Fassan,Raffaele Baffa,Mathew L. Thakur,Rick T. Owens,David J. McQuillan,Renato V. Iozzo.An Antimetastatic Role for Decorin in Breast Cancer[J]. The American Journal of Pathology . 2008 (3)
  • 2Fisher L W,Termine J D,Young M F.Deduced protein sequence of bone small proteoglycan I (biglycan) shows homology with proteoglycan II (decorin) and several nonconnective tissue proteins in a variety of species. Journal of Biological Chemistry . 1989
  • 3Krusius T,Ruoslahti E.Primary structure of an extracellular matrix proteoglycan core protein deduced from cloned cDNA. Proceedings of the National Academy of Sciences of the United States of America . 1986
  • 4Schaefer, Liliana,Iozzo, Renato V.Biological functions of the small leucine-rich proteoglycans: From genetics to signal transduction. Journal of Biological Chemistry . 2008
  • 5Bi Xiuli,Tong Chang,Dockendorff Ashley,Bancroft Laura,Gallagher Lindsay,Guzman Grace,Iozzo Renato V,Augenlicht Leonard H,Yang Wancai.Genetic deficiency of decorin causes intestinal tumor formation through disruption of intestinal cell maturation. Carcinogenesis . 2008
  • 6Tralh?o J Guilherme,Schaefer Liliana,Micegova Miroslava,Evaristo Cesar,Sch?nherr Elke,Kayal Samer,Veiga-Fernandes Henrique,Danel Claire,Iozzo Renato V,Kresse Hans,Lemarchand Patricia.In vivo selective and distant killing of cancer cells using adenovirus-mediated decorin gene transfer. The FASEB journal : official publication of the Federation of American Societies for Experimental Biology . 2003
  • 7Santra M,Eichstetter I,Iozzo R V.An anti-oncogenic role for decorin. Down-regulation of ErbB2 leads to growth suppression and cytodifferentiation of mammary carcinoma cells. Journal of Biological Chemistry . 2000
  • 8吕俊锋,杜珍武,张玉成,张桂珍.人核心蛋白聚糖基因靶向肺癌细胞特异性表达载体的构建及其表达[J].中国实验诊断学,2011,15(2):191-194. 被引量：2
  • 9王桂琴,兰晶,于培霞,邓志华.人核心蛋白聚糖真核表达载体的构建及其体内外抗瘤效应研究(英文)[J].山西医科大学学报,2011,42(9):697-703. 被引量：5
  • 10陈亮,熊雁,王子明,王爱民.核心蛋白聚糖抑制人软骨细胞MMP13基因表达的体外研究[J].第三军医大学学报,2012,34(8):719-722. 被引量：3

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中国老年学杂志
2011年 第5期

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