期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

利用绿色荧光蛋白筛选酵母突变株的新方法

A new method for screening mutant yeast strains with green fluorescent protein
下载PDF
导出
摘要 目的建立快速简便筛选酵母突变株的方法。方法利用同源重组技术,通过转座子上的GFP基因,从酵母基因组文库中,检测发生突变的菌株,从而筛选出酵母突变株。结果应用荧光显微镜观察SC-ura 选择平板筛选出的酵母,有绿色荧光发出的即为阳性菌落。结论利用同源重组技术,初步筛选出了所需的酵母突变株。 Objective To establish a fast and simple method for screening mutant yeast strains. Materials and Methods Homologous recombination technique was used to detect mutant yeast strains in yeast genomic library, with the green fluore- scent protein gene as the reporter gene in the transposon. Results The strains that emitted green fluorescence were isolated, indicating that the gfp gene was inserted into the yeast genome by homologous recombination. Conclusion This study established a useful method for functional genome study by homologous recombination technique, and provide an alternative for gene therapeutic drug development.
作者 胡莲美 朴英杰 何才姑 马文丽 崔东 郑文岭
机构地区 南方医科大学组织胚胎学教研室 南方医科大学分子生物学研究所 广州军区广州总医院医学实验科
出处 《第一军医大学学报》 CSCD 北大核心 2004年第10期1150-1152,共3页 Journal of First Military Medical University
基金 国家自然科学基金(39880044)~~
关键词 同源重组 转座子 绿色荧光蛋白 酵母 突变株 homologous recombination transposon green fluorescent protein yeast mutant strains
分类号 Q933 [生物学—微生物学]
  • 相关文献

参考文献11

  • 1Koller BH, Smithies O. Altering genes in animals by gene targeting[J]. Ann Rev Immunol, 1992, 10: 705-30.
  • 2Capecchi MR. Altering the genome by homologous recombineation[J]. Science, 1998, 244 (4910): 1288-92.
  • 3Ross-Macdonald P, Coelho PS, Roemer T, et al. Large-scale analysis of the yeast genome by transposon taging and gene disruption [J].Nature, 1999, 402(6760): 413-8.
  • 4Ross-macdonald AMY, Sheehan G. A multipurpose transposon system for analyzing protein production, localization, and function in Saccharomyces cerevisiae [ J ]. Proc Natl Acad Sci USA, 1997, 94:190-5.
  • 5Anuj K, Seema A, John A, et al. Subcellular localization of the yeast proteome[J]. Genes Dev, 2002, 16: 707-19.
  • 6萨姆布鲁克J 弗里奇EF 曼尼阿蒂斯T著.金冬雁 黎孟枫 译.分子克隆实验指南[M].第2版[M].北京:科学出版社,1992.456-494.
  • 7廖成,赵慕钧,李载平.小鼠一个新基因mLPTS的克隆、表达及亚细胞定位[J].Acta Genetica Sinica,2002,29(10):865-870. 被引量：6
  • 8Wood V, Gwilliam R, Rajandream M, et al. The genome sequence of Schizosaccharomyces pombe[ J ]. Nature, 2002, 415:871-80.
  • 9Giaever G, Chu AM, Ni L, et al. Functional profiling of the Sac charomyces cerevisiae genome [J]. Nature, 2002, 418 (6896):387-91.
  • 10Novak JE, Ross-Macdonald PB, Roeder GS. The budding yeast Msh4protein functions in chromosome synapsis and the regulation of crossover distribution[J]. Genetics, 2001, 158(3): 1013-25.

二级参考文献8

  • 1[1]Liao C, Zhao M J, Song H, Uchida K, Yokoyama K K, Li T P. Identification of the gene for a novel liver-related putative tumor suppressor at a high-frequency loss of heterozygosity region of chromosome 8p23 in human hepatocellular carcinoma. Hepatology, 2000, 32(4):721～727.
  • 2[2]Sambrook J, Fritsch E F, Maniatis T ed. Molecular cloning. Cold Spring Harbor Laborotory Press, 1989.
  • 3[3]Lippincott-Schwartz J. The uses of green fluorecent protein in mammalian cells. In: Chalfie M, Kain S ed. Green fluorescent protein: properties, applications and protocols. Wiley-Liss, 1998, 243～268.
  • 4[4]Horton P, Nakai K. Better prediction of protein cellular localization sites with the k nearest neighbors classifier. Ismb, 1997, 5: 147～152.
  • 5[5]Hicks G R, Raikhel N V. Protein import into the nucleus: an integrated view. Annu. Rev. Cell Dev. Biol., 1995, 11:155～188.
  • 6[6]Robbins J, Dilworth S M, Laskey R A, Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell, 1991, 64:615～623.
  • 7[7]Visintin R, Amon A. The nucleolus: the magician's hat for cell cycle tricks. Curr. Opin Cell Biol., 2000, 12: 372～377.
  • 8[8]Comai L. The nucleolus: a paradigm for cell proliferation and aging. Brazilian Journal of Medical and Biology Research, 1999, 32: 1473～1478.

共引文献26

第一军医大学学报
2004年 第10期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部