期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

CD146短发夹RNA真核表达载体的构建与鉴定

Construction and identification of the eukaryotic expression vector expressing short hairpin RNA targeting human CD146
下载PDF
导出
摘要 目的 CD146基因表达增高与涎腺腺样囊性癌(adenoid cystic carcinoma,ACC)嗜神经侵袭(perineural invasion,PNI)的临床症状相关。为进一步研究CD146基因的功能,构建人CD146短发夹RNA(short hairpin RNA,shRNA)真核表达载体并鉴定。方法根据Genebank中靶基因CD146序列,按照Tuschl设计原则,设计、合成编码其shRNA的互补寡核苷酸序列,克隆至线性化的pGenesil-1载体中并对重组质粒进行酶切分析及测序鉴定。结果经过限制性内切酶酶切分析和DNA测序鉴定,证实重组质粒pGe-CD146-shRNA的序列正确。结论成功构建了靶向人CD146基因的shRNA真核表达载体系统,为进一步研究CD146基因的功能奠定了实验基础。 Objective The overexpression of CD146 is reportedly associated with perineural invasion in salivary adenoid cystic carcinoma.This study was designed to construct and identify the eukaryotic expression vector expressing short hairpin RNA（shRNA） targeting human CD146.MethodsTwo pairs of shRNA were designed according to the human CD146 sequence in the Genebank（NM_006500） following the rules of Tuschl,and then synthesized and cloned into the linearized pGenesil-1 vector.The recombinant vector was confirmed by enzyme digestion analysis and DNA sequencing.ResultsThe inserted sequence of the recombinant pGe-CD146-shRNA vector was exactly demonstrated by double restrictive endonuclease digestion and sequence analysis.ConclusionThe pGe-CD146-shRNA eukaryotic expression vectors were successfully constructed and identified.
作者 陈伟 董震 蒋勇 苏寒 刘宝林 孙沫逸
机构地区 南京军区南京总医院口腔科 第四军医大学口腔医学院
出处 《医学研究生学报》 CAS 2011年第3期237-241,共5页 Journal of Medical Postgraduates
基金 国家自然科学基金(30772428) 南京军区南京总医院青年基金(2010Q009)
关键词 CD146 短发夹RNA 真核表达载体 CD146 Short hairpin RNA Eukaryotic expression vector
分类号 Q782 [生物学—分子生物学]
  • 相关文献

参考文献16

  • 1Fordice J,Kershaw C,el-Naggar A,et al.Adenoid cystic carcinoma of the head and neck.Predictors of morbidity and mortality[J].Arch Otolaryngol Head Neck Surg,1999,125(2):149-152.
  • 2Chen W,Zhang HL,Shao XJ,et al.Gene expression profile of salivary adenoid cystic carcinoma associated with perineural invasion[J].Tohoku J Exp Med,2007,212(3):319-334.
  • 3Johnson JP,Rothbacher U,Sers C.The progression associated antigen MUC18:A unique member of the immunoglobulin supergene family[J].Melanoma Res,1993,3(5):337-340.
  • 4Lehmann JM,Holzmann B,Breitbart EW,et al.Discrimination between benign and malignant cells of melanocytic lineage by two novel antigens,a glycoprotein with a molecular weight of 113000 and a protein with a molecular weight of 76000[J].Cancer Res,1987,47(3):841-845.
  • 5Lehmann JM,Riethmuller G,Johnson JP.MUC18,a marker of tumor progression in human melanoma,shows sequence similarity to the neural cell adhesion molecules of the immunoglobulin superfamily[J].Proc Natl Acad Sci USA,1989,86(24):9891-9895.
  • 6Shih IM,Elder DE,Speicher D,et al.Isolation and functional characterization of the A32 melanomaassociated antigens[J].Cancer Res,1994,54(9):2514-2520.
  • 7Bardin N,Frances V,Lesaule G,et al.Identification of the S-Endo 1 endothelial-associated antigen[J].Biochem Biophys Res Commun,1996,218(1):210-216.
  • 8Shih IM.The role of CD146(Mel-CAM) in biology and pathology[J].J Pathol,1999,89(1):4-11.
  • 9Yan X,Lin Y,Yang D,et al.A novel anti-CD146 monoclonal antibody,AA98,inhibits angiogenesis and tumor growth[J].Blood,2003,102(1):184-191.
  • 10Wu GJ,Varma VA,Wu MW,et al.Expression of a human cell adhesion molecule,MUC18,in prostate cancer cell lines and tissues[J].Prostate,2001,48(4):305-315.

二级参考文献20

  • 1葛棣,冯明祥,曾亮.表皮生长因子受体家族在食管癌中表达的临床研究[J].中国临床医学,2004,11(5):717-720. 被引量：11
  • 2Lee JC, Wang ST, Chow NH, et al. Investigation of the prognostic value of eoexpressed erbB family members for the survival of colorectal cancer patients after curative surgery[J]. Eur J Cancer, 2002,38 (8) : 1065-1071.
  • 3Graber HU, Friess H, Kaufmann B, et al. ErbB-4 mRNA expression is decreased in non-metastatic pancreatic cancer[J]. lnt J Cancer, 1999,84( 1 ) :24-27.
  • 4Abd E1-Rehim DM, Pinder SE, Paish CE, et al. Expression and co-expression of the members of the epidermal growth factor receptor (EGFR) family in invasive breast carcinoma [ J ]. Br J Cancer, 2004, 91 (8) :1532-1542.
  • 5Normanno N, Bianco C, Strlzzi L, et al. The ErbB receptors and their ligands in cancer: an overview [ J]. Curr Drug Targets, 2005,6(6) :243-257.
  • 6Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans [J]. Nature, 1998, 391(6669) :806-811.
  • 7Elbashir SM, Harborth J, Lendeckel W, et al. Duplexes of 21- nucleotide RNAs mediate RNA interference in cultured mammalian cells[J]. Nature, 2001, 411 (6836) :494-498.
  • 8Brummeikamp TR, Bernards R, Agami R. A system for stable expression of short interfering RNAs in mammalian cells[J]. Science, 2002, 296 (5567) :550-553.
  • 9von Lindern M, van Baal S, Wiegant J, et al. Can, a putative oncogene associated with myeloid leukemogenesis, may be activated by fusion of its 3 'half to different genes: characterization of the set gene[J]. Mol Cell Biol, 1992, 12(8):3346- 3355.
  • 10Karetsou Z, Martic G, Sflomos G, et al. The histone chaperone SET/TAF-Ibeta interacts functionally with the CREB-binding protein[J]. Biochem Biophys Res Commun, 2005, 335 ( 2 ) : 322- 327.

共引文献8

医学研究生学报
2011年 第3期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部