粟酒裂殖酵母基因组的Not I限制酶切图谱构建

Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome.

作者信息

Fan J B, Chikashige Y, Smith C L, Niwa O, Yanagida M, Cantor C R

机构信息

Department of Genetics and Development, College of Physicians and Surgeons, Columbia University, New York 10032.

出版信息

Nucleic Acids Res. 1989 Apr 11;17(7):2801-18. doi: 10.1093/nar/17.7.2801.

Pulsed field gel electrophoresis and large DNA technology were used to construct a Not I restriction map of the entire genome of the fission yeast Schizosaccharomyces pombe. There are 14 detectable Not I sites in S. pombe 972h: 9 sites on chromosome I and 5 sites on chromosome II, while no Not I sites were found on chromosome III. The 17 fragments (including intact chromosome III) generated by Not I digestion were resolved by PFG electrophoresis. These fragments ranged in size from 4.5 kb to approximately 3.5 Mb. Various strategies were applied in determining, efficiently, the order of the fragments on the chromosomes. The genomic size measured by adding all the fragments together is about 14 Mb and the sizes of the three chromosomes are I, 5.7 Mb, II, 4.6 to 4.7 Mb, and III, 3.5 Mb. These are generally somewhat smaller than estimated previously.

采用脉冲场凝胶电泳和大片段DNA技术构建了裂殖酵母粟酒裂殖酵母全基因组的Not I限制性酶切图谱。在粟酒裂殖酵母972h中有14个可检测到的Not I位点：染色体I上有9个位点，染色体II上有5个位点，而在染色体III上未发现Not I位点。Not I酶切产生的17个片段（包括完整的染色体III）通过脉冲场凝胶电泳进行分离。这些片段大小从4.5 kb到约3.5 Mb不等。采用了各种策略来高效确定染色体上片段的顺序。将所有片段大小相加测得的基因组大小约为14 Mb，三条染色体的大小分别为：染色体I，5.7 Mb；染色体II，4.6至4.7 Mb；染色体III，3.5 Mb。这些数值总体上比之前估计的略小。