Neitz M, Carbon J
Mol Cell Biol. 1985 Nov;5(11):2887-93. doi: 10.1128/mcb.5.11.2887-2893.1985.
A functional centromere located on a small DNA restriction fragment from Saccharomyces cerevisiae was identified as CEN14 by integrating centromere-adjacent DNA plus the URA3 gene by homologous recombination into the yeast genome and then by localizing the URA3 gene to chromosome XIV by standard tetrad analysis. DNA sequence analysis revealed that CEN14 possesses sequences (elements I, II, and III) that are characteristic of other yeast centromeres. Mitotic and meiotic analyses indicated that the CEN14 function resides on a 259-base-pair (bp) RsaI-EcoRV restriction fragment, containing sequences that extend only 27 bp to the right of the element I to III region. In conjunction with previous findings on CEN3 and CEN11, these results indicate that the specific DNA sequences required in cis for yeast centromere function are contained within a region about 150 bp in length.
通过同源重组将着丝粒相邻DNA加上URA3基因整合到酵母基因组中,然后通过标准四分体分析将URA3基因定位到第十四号染色体上,从而鉴定出酿酒酵母一个小DNA限制片段上的功能性着丝粒CEN14。DNA序列分析表明,CEN14拥有其他酵母着丝粒所特有的序列(元件I、II和III)。有丝分裂和减数分裂分析表明,CEN14的功能位于一个259碱基对(bp)的RsaI - EcoRV限制片段上,该片段包含的序列仅从元件I至III区域向右延伸27 bp。结合之前关于CEN3和CEN11的研究结果,这些结果表明酵母着丝粒功能顺式所需的特定DNA序列包含在一个长度约为150 bp的区域内。