文章基本信息

标题：Konstruksi Strain Haploid Saccharomyces cerevisiae Pembawa Kromosom XII yang Terpotong pada Sisi Kanan Lokus rDNA
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作者：TRIASTUTI RAHAYU ; DONNY WIDIANTO ; TRIWIBOWO YUWONO 等
期刊名称：Bioteknologi
印刷版ISSN：0216-6887
出版年度：2005
卷号：2
期号：1
页码：9-13
DOI：10.13057/biotek/c020102
出版社：UNS Solo
摘要：Saccharomyces cerevisiae has a great potential for fermentation industryand molecular biology research. Many works have been conducted toincrease S. cerevisiae growth rate. The objective of this research was toobtain a rDNA fragment of chromosome XII of S. cerevisiae with splitting atright region of rDNA locus. Chromosome XII was splitted by using achromosome-splitting vector (pDW49) that carried a cloned DNA fragmentof rDNA locus. An E. coli strain DH5α was used as a host for pDW49amplification. The pDW49 plasmid was digested with BamHI to eliminatethe HIS3 DNA stuffer. The larger BamHI fragment was isolated by agarosegel electrophoresis. The BamHI fragment was subsequently self-ligated,resulting in the Tr ends being joined head-to-head. The recircularized DNAmolecule was linearized by cleavage at the homologous sequence (at theright region of rDNA locus) using BglII. The linearized DNA molecule wasintroduced into S. cerevisiae strain W303-1A by lithium acetate method.Confirmation of chromosome XII splitting was analyzed by PFGE (PulsedField Gel Electrophoresis) for 24 h with switching interval of 45 secfollowed by 6 h run with switching interval of 15 sec. The result of PFGEshowed an additional chromosomal band (611 kbp), suggesting that thechromosome XII has been splitted
关键词：Saccharomyces cerevisiae; Chromosome XII and rDNA.