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mRNA差别显示技术分离草菇低温诱导基因*
引用本文:陈明杰,谭琦,曹晖,王南,黄为一,潘迎捷.mRNA差别显示技术分离草菇低温诱导基因*[J].菌物学报,2001,20(3).
作者姓名:陈明杰  谭琦  曹晖  王南  黄为一  潘迎捷
作者单位:1 南京农业大学资环学院 2 农业部食用菌遗传育种重点开放实验室,上海市农业遗传育种重点开放实验室,上海市农业科学院食用菌研究所
摘    要:本文报道采用改良的mRNA差别筛选法,通过选用3'锚定引物和18-25碱基的PCR随机引物组合,以琼脂糖凝胶电泳,EB显色替代聚丙烯酰胺凝胶电泳和放射自显影,对低温处理草菇(Volvariella volvacea)及正常草菇基因表达进行筛选、分析,结合RNA斑点杂交技术加以验证,分离得到70个草菇低温诱导DNA片段。

关 键 词:草菇  mRNA差别筛选法  低温诱导

ISOLATION OF COLD INDUCED GENE FORM VOLVARIELLA VOLVACEA BY mRNA DIFFERENTIAL DISPLAY
Abstract:Modified mRNA differential display technique was used in this study. With cDNA as template, 3' anchor primer and 18-25 random primers were used for PCR amplification. Agarose gel electrophoresis and ethidium bromide staining were used instead of PAGE and autoradiograph for detection. 70 cold induced genes of Volvariella volvacea were obtained by this modified mRNA differential display technique and cold specific DNAs were identified by RNA dot blot hybridization.
Keywords:Volvariella volvacea  mRNA differential display cold induced gene
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