...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Cloning and characterization of LOS1, a Saccharomyces cerevisiae gene that affects tRNA splicing.
【24h】

Cloning and characterization of LOS1, a Saccharomyces cerevisiae gene that affects tRNA splicing.

机译：LOS1的克隆和鉴定，这是一种酿酒酵母基因，可影响tRNA的剪接。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Saccharomyces cerevisiae strains carrying los1-1 mutations are defective in tRNA processing; at 37 degrees C, such strains accumulate tRNA precursors which have mature 5' and 3' ends but contain intervening sequences. Strains bearing los1-1 and an intron-containing ochre-suppressing tRNA gene, SUP4(0), also fail to suppress the ochre mutations ade2-1(0) and can1-100(0) at 34 degrees C. To understand the role of the LOS1 product in tRNA splicing, we initiated a molecular study of the LOS1 gene. Two plasmids, YEpLOS1 and YCpLOS1, that complement the los1-1 phenotype were isolated from the YEp24 and YCp50 libraries, respectively. YEpLOS1 and YCpLOS1 had overlapping restriction maps, indicating that the DNA in the overlapping segment could complement los1-1 when present in multiple or single copy. Integration of plasmid DNA at the LOS1 locus confirmed that these clones contained authentic LOS1 sequences. Southern analyses showed that LOS1 is a single copy gene. The locations of the LOS1 gene within YEpLOS1 and YCpLOS1 were determined by deletion and gamma-delta mapping. Two genomic disruptions of the LOS1 gene were constructed, i.e., an insertion of a 1.2-kilobase fragment carrying the yeast URA3 gene, los1::URA3, and a 2.4-kilobase deletion from the LOS1 gene, los1-delta V. Disruption or deletion of most of the LOS1 gene was not lethal; cells carrying the disrupted los1 alleles were viable and had phenotypes similar to those of cells carrying the los1-1 allele. Thus, it appears that the los1 gene product expedites tRNA splicing at elevated temperatures but is not essential for this process.
机译：带有los1-1突变的酿酒酵母菌株在tRNA加工中存在缺陷。在37℃下，这些菌株积累了具有成熟的5'和3'末端但包含插入序列的tRNA前体。带有los1-1和含内含子的抑制tRNA基因SUP4（0）的菌株在34摄氏度时也无法抑制茶色突变ade2-1（0）和can1-100（0）。在tRNA剪接过程中对LOS1产品进行了研究，我们启动了LOS1基因的分子研究。与los1-1表型互补的两个质粒YEpLOS1和YCpLOS1分别从YEp24和YCp50文库中分离出来。 YEpLOS1和YCpLOS1具有重叠的限制性图谱，表明重叠片段中的DNA以多拷贝或单拷贝形式存在时可以与los1-1互补。质粒DNA在LOS1位点的整合证实了这些克隆包含真实的LOS1序列。 Southern分析表明LOS1是单拷贝基因。 YEpLOS1和YCpLOS1中LOS1基因的位置是通过缺失和伽马-δ作图确定的。构建了LOS1基因的两个基因组破坏，即插入了带有酵母URA3基因los1.2 :: URA3的1.2碱基碱基片段，以及从LOS1基因los1-delta V中删除了2.4碱基碱基。大多数LOS1基因不是致命的。携带破坏的los1等位基因的细胞是可行的，其表型与携带los1-1等位基因的细胞相似。因此，似乎los1基因产物在升高的温度下加速了tRNA剪接，但对于该过程不是必需的。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号