期刊文献+

原位杂交法检测hsp mRNA在猪组织细胞中的分布

Detection of hsp mRNA in the tissues of pigs by in situ hybridization
下载PDF
导出
摘要 根据hsp70和hsp90 mRNA基因序列,设计并合成引物,将PCR扩增的基因片段克隆到pGEM-T载体,重组质粒经筛选、鉴定,利用体外转录系统,制作地高辛标记的RNA探针,建立检测hsps mRNA原位杂交方法。利用自建的原位杂交方法,对hsp70、hsp90 mRNA进行组织细胞定位研究。结果显示:hsp70 mRNA和hsp90 mRNA广泛分布于各组织细胞中,尤其在细胞核中相对集中,在肾小管上皮细胞胞浆中也呈强阳性着色。 According to the gene sequences of hsp70 and hsp90 mRNA, the primers were designed and synthesized. The fragments generated by RT-PCR were cloned into the pGEM-T vector and the recombinant plasmids were identified by PCR amplification. Digoxigenin (DIG) labeled RNA probes were prepared by transcription in vitro in order to establish ISH (in situ hybridization) method. The distributions of hsp70 mRNA and hsp90 mRNA in the tissues of pigs were examined by means of ISH method. Hsp70 mRNA and hsp90 mRNA were found in most of the detected tissues, preferentially in the nuclei and also in the cytoplasm of renal tubular epithelial cells.
出处 《畜牧与兽医》 北大核心 2008年第3期18-21,共4页 Animal Husbandry & Veterinary Medicine
基金 国家自然科学基金(30430420,39770569,30170682) 聊城大学博士科研启动基金(31805)
关键词 原位杂交 HSP MRNA 体外转录 in situ hybridization hsp mRNA transcription in vitro pigs
  • 相关文献

参考文献16

  • 1Arrigo A P,Landry J.Expression and function of the low-molecular weight heat shock proteins.In:Morimoto R I.Tissieres A.Georgopoulos C(eds.).Biology of Heat Shock Proteins and Molecular Chaperons.Cold Spring Harbor Laboratory Press,1994:335-373.
  • 2杜立银,田文儒.热应激对小鼠附植前早期胚胎HSP70的诱导表达[J].畜牧与兽医,2006,38(10):9-11. 被引量:2
  • 3Shyu W C,Kao M C,Chou W Y,et al.Heat shock modulates prion protein expression in human NT-2 Cells[J].Molecular Neuroscience,2000,2(4):771-774.
  • 4Saavedra C A,Hammell C M,Heath C V,et al.Yeast heat shock mRNAs are exported through a distinct pathway defined by Riplp[J].Genes and Development,1997,11:2845-2856.
  • 5Vainberg I E,Dower K,Rosbash M.Nuclear export of heat shock and non-heat-shock mRNA Occurs via similar pathways[J].Molecular and Cellular Biology,2000,20:3996-4005.
  • 6Saavedra C,Tung K S,Amberg D C,et al.Regulation of mRNA export inresponse to stress in Saccharomyces cerevisiae[J].Genes and Development,1996,10:1608-1620.
  • 7Stutz F,Izaurralde E,Mattaj I W,et al.A role for nueleoporin FG repeat domains inexport of human immunodeficiency virus type 1Bey protein and RNA from the nucleus[J].Molecular and Cellular Biology,1996,16:7144-7150.
  • 8俞峻,马康涛.分子伴侣的多重功能[J].生物化学与生物物理进展,1998,25(2):106-110. 被引量:36
  • 9de Benedetti A,Bnglioni C.Translational regulation of the synthesis of a major heat shock protein in HeLa cells[J].The Journal of Bio-logical Chemistry,1986,261(33):15800-15804.
  • 10LI Yu-bao,BAO En-dong,WANG Zhi-liang,ZHAO Ru-qian.Detection of HSP mRNA Transcription in Transport Stressed Pigs by Fluorescence Quantitative RT-PCR[J].Agricultural Sciences in China,2007,6(3):361-367. 被引量:6

二级参考文献30

  • 1李玉保,付旭彬,鲍恩东.热休克蛋白研究进展[J].畜牧与兽医,2004,36(11):38-40. 被引量:12
  • 2钟文英,普雄明.热休克蛋白的分子生物学研究进展[J].医学综述,2005,11(2):148-150. 被引量:44
  • 3Arrigo A P,Landry J.Expression and function of the low-molecular weight heat shock proteins.In:Morimoto R I.Tissieres A,Georgopoulos C (eds.).Biology of Heat Shock Proteins and Molecular Chaperons.Cold Spring Harbor Laboratory Press,1994:335-373.
  • 4Garrido C,Ottavi P,Fromentin A,Hammann A,Arrigo A P,Chauffert B,Mehlen P.HSP27 as a mediator of confluence-dependent resistance to cell death induced by anticancer drugs.Cancer Research,1997,57:2 661-2 667.
  • 5Shyu W C,Kao M C,Chou W Y,Hsu Y D,Soong B W.Heat shock modulates prion protein expression in human NT-2 Cells.Molecular Neuroscience,2000,2:771-774.
  • 6Saavedra C A,Hammell C M,Heath C V,Cole C N.Yeast heat shock mRNAs are exported through a distinct pathway defined by Riplp.Genes and Development,1997,11:2845-2856.
  • 7Vainberg I E,Dower K,Rosbash M.Nuclear export of heat shock and non-heat-shock mRNA occurs via similar pathways.Molecular and Cellular Biology,2000,20:3996-4005.
  • 8Saavedra C,Tung K S,Amberg D C,Hopper A K,Cole C N.Regulation of mRNA export inresponse to stress in Saccharomyces cerevisiae.Genes and Development,1996,10:1608-1620.
  • 9Stutz F,Izaurralde E,Mattaj I W,Rosbash M.A role for nucleoporin FG repeat domains inexport of human immunodeficiency virus type 1 Rev protein and RNA from the nucleus.Molecular and Cellular Biology,1996,16:7144-7150.
  • 10de Benedetti A,Baglioni C.Translational regulation of the synthesis of a major heat shock protein in HeLa cells.The Journal of Biological Chemistry,1986,261:15800-15804.

共引文献66

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