外源核酸促核辐射鼠肠腺细胞修复的基因分析
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“211”工程基金资助(98X207).


Analysis of Genes Associated with Exogenous Nucleic Acids Improving the Repair of Intestinal Epithelium After γ Irradiation in Mice
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This study was supported by grants from 211 project fund (98X207).

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    摘要:

    初步探讨外源核酸促电离辐射损伤的小鼠肠腺细胞修复的分子机理.建立BALB/c小鼠电离辐射后给予外源RNA与生理盐水治疗的6 h、12 h、24 h、4 d和8 d的模型,采集空肠组织标本后采用消减杂交基础上的LD-PCR技术,获取与受照小鼠肠腺损伤修复相关的基因克隆,对其进行全自动序列分析与GenBank检索.6 h治疗组同源性较高的序列主要为:热休克蛋白mRNA、Nmi mRNA、Dutt1蛋白mRNA、Na,K-ATPase γ亚单位mRNA等;12 h治疗组同源性较高的序列为:碱性磷酸酶mRNA、碱性磷酸酶2、glkA基因、单链复制着丝粒基因等;24 h治疗组同源性较高的序列为:抗CEA单链抗体重链可变区基因、抗DNA重链可变区基因、Ig kappa链mRNA等;4 d治疗组同源性较高的序列为:双特异性磷酸酶、端粒酶相关蛋白家族mRNA、β-GABA转运基因、紧张激活蛋白mRNA、FK506结合蛋白、Ca2+/Ca2+调蛋白依赖性基因等;8 d治疗组同源性较高的序列为:免疫球蛋白可变区基因、鼠免疫球蛋白DNA、易弯曲肽DNA、tsr glkA基因、修复蛋白A等.新发现的18个基因片段递交给GeneBank,接受号为AF240164-AF240181.结果表明:外源核酸促电离辐射损伤的小鼠肠腺修复的机理可能与一些基因、蛋白质的异常表达有关,与免疫系统的作用可能有关.

    Abstract:

    In order to explore the molecular mechanism of exogenous nucleic acids improving repair of irradiation-damaged intestinal epithelium, 45 mice being irradiated by γ ray were treated with 40 μg small intestinal RNA as test group, whose small intestinal specimens were collected respectively at 6 h,12 h,24 h,4 d and 8 d after treatment; 40 mice being irradiated by γ ray were treated with physiological saline as control group, whose small intestinal specimens were collected at the same interval time. Then fragments of genes expressed in test group higher than those in control group, were obtained by using LD-PCR based on subtractive hybridization. After that, these gene fragments were cloned into T vectors,and were sequenced. Obtained sequences were searched for GenBank.90 clones associated with repair of irradiation -damaged crypt cells were obtained.In test group of 6 h, higher similar sequences mainly were as follows: mRNA for heat shock protein, Nmi mRNA, Dutt1 protein, mRNA for Na,K-ATPase gamma subunit,mRNA for surface glycoprotein,Zinc finger type transcript factor,porcine growth hormone-releasing hormone gene,Homo sapiens dual specificity phosphatase,etc. In test group of 12 h, higher similar sequences were as follows: alkaline phosphatase mRNA,alkaline phosphatase 2,glkA gene, single stranded replicative centromeric gene,Homo sapiens DMBT1 candidate tumor gene, tRNA -Met gene,mouse Ig unrearranged transcribed H-chain,thyroxine-binding globulin gene,alpha-2-plasmin inhibitor gene, etc;In test group of 24 h, higher similar sequences were as follows: anti-CEA ScFv antibody heavy chain vary region,anti-DNA antibody Ig heavy chain, mRNA for Ig kappa chain region,anti-BONT/A Hc ScFv antibody heavy chain vary region, mRNA for ScFv collagenase heavy chain vary region, AE0199 immunoglobulin heavy chain,mouse Ig gamma-chain,Ig rearranged gamma-chain mRNA,anti-NP antibody IgH,mRNA for arginine/serine kinase,dual specificity phosphatase,family mRNA telomerase-associated protein,anti-human erB-2 region,BMP-4 gene,etc; In test group of 4 d, higher similar sequences were as follows: mRNA for sodium channel,tazarotene-induced gene,betaine-GABA transporter gene,homobox protein Xgbx-2 mRNA,mRNA for stress-activated protein,FK506 binding protein,calium /calmodulin dependent gene,PEST phosphatase interactin gene,haptoglobin mRNA, etc;In test group of 8 d, higher similar sequences were as follows: Ig Mu variable region mRNA,Mus musculus Ig K chain mRNA V-region, mRNA for Hox1b protein,Mus musculus neutroactin mRNA, rat alkaline phosphatase mRNA,Human mRNA for XP-C repair complementing protein,human alpha-2-plasmin inhibitor gene,mRNA for CCAT binding factor,mouse active H-chain VJ region,etc. Eighteen were new sequences,whose function were unclear. Ninety clones were obtained to be associated with repair of damaged mice intestinal gland cells caused by γ ray and treated by small intestinal RNA.Repair of damaged intestinal gland cells treated by exogenous nucleic acids may be associated with hsp,Nmi,Dutt1,alkaline phosphatase genes and eighteen new sequences.

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崔大祥,曾桂英,王枫,田芙蓉,郭晏海,徐俊荣,闫小君,任东清,苏成芝.外源核酸促核辐射鼠肠腺细胞修复的基因分析[J].生物化学与生物物理进展,2001,28(3):353-357

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  • 收稿日期:2000-05-08
  • 最后修改日期:2000-07-07
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