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应用生态学报 ›› 2001, Vol. ›› Issue (2): 269-271.

• 研究论文 • 上一篇    下一篇

卤代烷烃脱卤酶基因在拟南芥中反式失活系统的建立

郝林1, 曹军2, 冯树2   

  1. 1. 沈阳师范学院生物系, 沈阳110034;
    2. 中国科学院沈阳应用生态研究所, 沈阳110016
  • 收稿日期:2000-02-18 修回日期:2000-04-11 出版日期:2001-03-25
  • 通讯作者: 郝林,男,1963年生,中国科学院沈阳应用生态研究所在读博士研究生,导师张成刚研究员和张忠泽研究员,研究方向为分子生物学和微生物工程.在国内外学术刊物上发表研究论文30余篇.
  • 基金资助:
    中国科学院知识创新工程资助项目(KZCX2401)

Establishment of trans-inactivation system for haloalkane dehalogenase gene in Arabidopsis thaliana

HAO Lin1, CAO Jun2, FENG Shu 2   

  1. 1. Biological Department, Shenyang Teacher’s College, Shenyang 110034;
    2. Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016
  • Received:2000-02-18 Revised:2000-04-11 Online:2001-03-25

摘要: 以细菌Xanthobacterautotrophicus卤代烷烃脱卤酶基因为遗传负选择标记,建立了该基因在拟南芥中反式失活的实验系统.在卤代烷烃脱卤酶转基因拟南芥中,有1株表现为转基因失活,离体核run-off转录实验表明为基因转录后沉默(这里特指沉默位点).用这一转基因沉默植株与同源转基因高效表达植株(这里特指同源转基因位点)杂交,结果96%的F1代植株表现为同源转基因反式失活.将F1代植株自交,使部分沉默位点与反式失活的同源转基因位点分离,结果200株子代中有42株表现DhlA活性,158株无DhlA活性,即dhlA沉默植株与表达植株之比为3.76∶1,表明沉默位点是以孟德尔显性因子方式使同源转基因位点反式失活的.

Abstract: With haloalkane dehalogenase gene (dhlA) of Xanthobacter autothophicus as a negative selection marker,a novel trans-inactivation system for transgenes in plants was developed.After the A rabidopsis plants were transformed with dhlA, one plant was shown dhlA post-transcriptional silencing by nuclear run-off transcriptional analysis.The A rabidopsis plants containing silencing locus were crossed to those efficiently expressed dhlA,referred as a homologous transgenic locus,and 96% of F1 plants became the homologous dhlA inactivated in trans. When the F1 plants were selfed,and the silencing locus and the homologous locus were separated in some progenies,42 of the 200 progenies exhibited DhlA activity,and 158 no DhlA activity, and the proportion of the DhlA-expressing plants to the dhlA-silencing ones was 3.76 to 1,which suggested that the silencing locus inactivated in trans the homologous transgenic locus as a Mendelian dominant factor.

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