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尹元琴, 任常山. 人神经生长因子cDNA的体外扩增[J]. 中国公共卫生, 2001, 17(6): 495-496. DOI: 10.11847/zgggws2001-17-06-11
引用本文: 尹元琴, 任常山. 人神经生长因子cDNA的体外扩增[J]. 中国公共卫生, 2001, 17(6): 495-496. DOI: 10.11847/zgggws2001-17-06-11
YIN Yuan-qin, REN Chang-shan. Amplification of Human Nerve Growth Factor cDNA in Vitro[J]. Chinese Journal of Public Health, 2001, 17(6): 495-496. DOI: 10.11847/zgggws2001-17-06-11
Citation: YIN Yuan-qin, REN Chang-shan. Amplification of Human Nerve Growth Factor cDNA in Vitro[J]. Chinese Journal of Public Health, 2001, 17(6): 495-496. DOI: 10.11847/zgggws2001-17-06-11

人神经生长因子cDNA的体外扩增

Amplification of Human Nerve Growth Factor cDNA in Vitro

  • 摘要: 目的 提取人胎盘组织中RNA,以其中的mRNA为模板,扩增人βNGFcDNA,并测定核苷酸序列,为进一步克隆及表达βNGFcDNA奠定基础.方法 采用异硫氢酸胍法提取RNA;采用RT-PCR技术扩增人βNGFcDNA;Sanger双脱氧链终止法测定核苷酸序列.结果 从人胎盘组织中提取RNA,以其中的mRNA为模板,扩增出人βNGFcDNA,经琼脂糖凝胶电泳鉴定为目的片段,序列测定证实与实际序列相符.结论 以人胎盘组织中的mRNA为模板,运用RT-PCR技术扩增βNGFcDNA,扩增时在5'端以限制性内切酶酶切位点加以修饰,以便于βNGF基因的克隆及表达.

     

    Abstract: Objective RNA was extracted from human placenta.Human βNGFcDNA was amplified by using mRNA as template and then nucleotide sequence was deter mined.All of this work is the basis of further study of cloning and expression of βNGFcDNA.Methods RNA was extracted by guanidine isothiocyanate method;human βNGFcDNA was amplified by RT-PCR technology;nucleoside sequence was identified using Sanger double deoxgen end method.Results RNA was extracted from the tissue of human placenta;human βNGFcDNA was amplified,mRNA of RNA served as template.The segment was identified as aimed DNA by agarose electrophoresis,and necleotide sequence accorded to actual sequence.Conclusion Human βNGFcDNA was amplified by RT-PCR technology using mRNA from human placenta as template and cDNA 5' end was modified with cutting-site of restriction endonuclease,which is convenient for the cloning and expression of βNGF gene.

     

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