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一种快速·有效分离T-DNA插入位点的侧翼序列的方法——DW-ACP-PCR技术

A Rapid and Efficient Method of Isolating the Flanking Sequence of T-DNA Inserted Site-DW-ACP-PCR Technique
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摘要 DW-ACP-PCR(DNAWalking-Annealing control primer-PCR)是一种分离已知DNA序列的侧翼未知序列的新技术。该技术通过3个嵌套的特异引物分别和ACPC复性控制引物组合,进行3步连续的PCR反应,所有反应可以在1 d内完成。ACP的特殊结构能有效地控制非特异扩增。用DW-ACP-PCR技术扩增7个稻瘟菌T-DNA插入突变体的T-DNA插入位点的侧翼序列,均获得了特异目标片段。 DW-ACP-PCR(DNA Walking Annealing control primer PCR) was a new technique of isolating the unknown flanking sequences in known DNA sequence,in which 3 nested specific primers were resp.combined with ACPC renaturation control primers for 3 step continuous PCR reactions.All the reactions needed only one day to be accomplished.The unique structure of ACP could control nonspecific amplification effectively.It was introduced that DW-ACP-PCR technique was used to amplify the flanking sequences of T-DNA inserted site of 7 T-DNA inserted mutants of Magnaporthe grisea.
出处 《安徽农业科学》 CAS 北大核心 2007年第28期8817-8818,8841,共3页 Journal of Anhui Agricultural Sciences
基金 国家自然科学基金(30260046)
关键词 DW-ACP-PCR T-DNA定位 侧翼序列 稻瘟菌 DW-ACP-PCR T-DNA location Flanking sequences Magnaporthe grisea
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  • 1Jong-Seong J, Gynheung An. Gene tagging in rice: A hight throughput system for functional genomies. Plant Sci, 2001,161:211-219.
  • 2Baraka A, Gallois P, Raynal M, et al. The distribution of T-DNA in the genomes of transgenic Arabidopsis and rice.FEBS Lett, 2000, 471:161-164.
  • 3Sessions A, Burke E, Presting G, et al. A High-throughput Arabidopsis reverse genetics system. Plant Cell, 2002, 14:2985-2994.
  • 4Perucho M, Hanahan D, Leah L, et al. Isolation of the chicken thymidine kinase gene by plasmid rescue. Nature, 1980, 285:207-210.
  • 5Kiessling U, Platzer M, Sreauss M. Plasmid resuce, a tool for reproducible recovery of genes from transfected mammalian cell? Mol Genet Genom, 1984, 193:513-519.
  • 6Nakazawa M, Yabe N, Ichikawa T, et al. DFL1, an auxin-responsive GH3 gene homologue, negatively regulatess hoot cell elongation and lateral root formation, and positively regulates the light response of hypocotyl length.Plant J, 2001, 25:213-221.
  • 7Migeon J C, Garfinkel M S, Edgar B A. Cloning and characterization of peter pan, a novel Drosophila gene required for larval growth. Mol Biol Cell, 1999, 10:1733-1744.
  • 8Mathur J, Szabados L, Sabine S, et al. Gene identification with sequenced T-DNA tags generated by transformation of Arabidopsis cell suspension. Plant J, 1998, 13:707-716.
  • 9Liu Y-G, Mitsukawa N, Oosumi T, et al. Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J, 1995, 8:457--463.
  • 10Zabarovsky E R, Weinberg G. High efficiency electroporation of ligated DNA into bacteria. Nucl Acids Res,1990, 18:5912.

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